United States Environmental

Protection Agency

Office of Water

Washington, DC 20460

EPA-822-R-03-013

May 2003

Methods for evaluating wetland condition

#14 Wetland Biological Assessment

Case Studies
United States Environmental

Protection Agency

Office of Water

Washington, DC 20460

EPA-822-R-03-013

May 2003

Methods for evaluating wetland condition

#14 Wetland Biological Assessment

Case Studies

Edited By

Maine Department of Environmental Protection

Thomas J. Danielson

Connecticut Department of Environmental Protection

Douglas G. Hoskins

Prepared jointly by:

The U.S. Environmental Protection Agency

Health and Ecological Criteria Division (Office of Science and Technology)

and

Wetlands Division (Office of Wetlands, Oceans, and Watersheds)
United States Environmental

Protection Agency

Office of Water

Washington, DC 20460

EPA-822-R-03-013

May 2003

Notice

The material in this document has been subjected to U.S. Environmental Protection Agency (EPA)

technical review and has been approved for publication as an EPA document. The information

contained herein is offered to the reader as a review of the “state of the science” concerning wetland

bioassessment and nutrient enrichment and is not intended to be prescriptive guidance or firm advice.

Mention of trade names, products or services does not convey, and should not be interpreted as

conveying official EPA approval, endorsement, or recommendation.

Appropriate Citation

U.S. EPA. 2003. Methods for Evaluating Wetland Condition: Wetland Biological Assessment

Case Studies. Office of Water, U.S. Environmental Protection Agency, Washington, DC.

EPA-822-R-03-013.

Acknowledgments

EPA acknowledges the contributions of the following people in the writing of this module: The members

of BAWWG—without their hard work, this report would not have possible. Leigh Dunkelberger

provided invaluable assistance during her internship with the U.S. EPA Wetlands Division.

This entire document can be downloaded from the following U.S. EPA websites:

http://www.epa.gov/ost/standards

http://www.epa.gov/owow/wetlands/bawwg

ii
iii

Contents

Foreword .................................................................................................................. vii

List of “Methods for Evaluating Wetland Condition” Modules ............. ix

Florida: Monitoring Across a Nutrient Gradient in the Everglades ..... 1

Florida: Development of a Biological Approach for

Assessing Florida Wetland Integrity ............................................................... 7

Maine: Developing a Statewide Biological Monitoring

and Assessment Program for Freshwater Wetlands................................ 14

Maryland: Developing an IBI Assessment for Restored

Wetlands in the Mid-Atlantic States ............................................................... 18

Massachusetts: Use of Multimetric Indices to Examine Ecological

Integrity of Salt Marsh Wetlands in Cape Cod ..........................................22

Massachusetts: Involving Volunteers in Examining the

Ecological Integrity of Coastal Wetlands in Cape Cod,

Massachusetts .......................................................................................................30

Michigan: Bioassessment Procedures and Baseline Reference

Data for Great Lakes Coastal Marshes and Inland Forested

Wetlands in Michigan ...........................................................................................32

Minnesota: Developing Wetland Biocriteria ...............................................37

Minnesota: Dakota County Wetland Health Evaluation Project ........44

Minnesota: University of Minnesota’s IBI Development ............................ 47

Montana: Developing Wetland Bioassessment Protocols To

Support Aquatic Life Beneficial Use-Support Determinations ...............49

North Dakota: Wetland Bioassessment Protocols for Making

Aquatic Life Beneficial Use-Support Determinations ................................59

Ohio: Developing IBI Assessment Methods for Water Quality

Standards and Regulatory Decisions .............................................................63

Oregon: Simultaneous Development, Calibration, and Testing

of Hydrogeomorphic-Based (HGM) Assessment Procedures and

Biological Assessment Procedures ...............................................................72
iv

Pennsylvania: Assessing Wetland Condition at a Watershed

Scale Using Hydrogeomorphic Models and Measures of

Biological Integrity ..............................................................................................75

Vermont: Classification, Biological Characterization, and

Biometric Development for Northern White Cedar Swamps

and Vernal Pools ..................................................................................................77

Washington: King County Wetland-Breeding Amphibian

Monitoring Program .............................................................................................80

Wisconsin: Developing Biological Indexes for Wisconsin’s

 Palustrine Wetlands ...........................................................................................82

Wisconsin: Refinement and Expansion of the Wisconsin Wetland

Biological Index for Assessment of Depressional Palustrine

Wetlands..................................................................................................................86

References .............................................................................................................. 91

Glossary...................................................................................................................93

List of Figures

Figure 1: Area of interest within Everglades protection area ............ 1

Figure 2: WCA 2A monitoring sites along the phosphorus gradient .. 2

Figure 3: Change point analyses of Eleocharis frequency of

occurrence and biomass data along the SFWMD

transects.............................................................................................. 5

Figure 4: Results of change point analyses performed on

median total percentage of pollution-sensitive

(literature determined) periphyton taxa .................................... 5

Figure 5: Results of change point analyses on

median Florida index values ............................................................ 6

Figure 6: Florida’s wetland regions and field site locations ............... 9

Figure 7: Relationships between wetland ecological condition

and land use .......................................................................................27

Figure 8: Two sets of multimetric goals for evaluating

restoration ........................................................................................28
v

Figure 9: Locations of smaller depressional wetlands

sampled in 1995 ................................................................................39

Figure 10: Locations of the 44 large depressional wetlands

sampled in 1999 ................................................................................39

Figure 11: Montana ecoregions and pilot watersheds ............................ 51

Figure 12: Ecoregions and sampling locations by wetland class .......52

Figure 13: Diatom data related to abiotic factors using

Detrended Canonical Correspondence Analysis.................54

Figure 14: Clusters of wetlands based on diatom data ..........................54

Figure 15: Macroinvertebrate index scores for wetlands

in several classes ...........................................................................57

Figure 16: Level IV ecoregions of North Dakota ....................................... 61

Figure 17: Ecological regions of Ohio and Indiana ..................................64

List of Plates

Plate 1: Cypress wetland .............................................................................. 12

Plate 2: Macroinvertebrate sampling ....................................................... 13

Plate 3: Maine’s stovepipe sampler and sieve bucket ............................ 16

Plate 4: Picking macroinvertebrates from multihabitat sample ....... 16

Plate 5: Cape Cod, MA, salt marsh plant survey ....................................29

Plate 6: Cape Cod, MA, salt marsh nekton: Watch your fingers!

(blue crab, Callinectes sapidus) .................................................29

Plate 7: Impaired wetland ..............................................................................66

Plate 8: Least-disturbed wetland ...............................................................66
vi

List of Tables

Table 1. Scoring Criteria for 10 Invertebrate Metrics for IBI

for Large Depressional Wetlands ............................................42

Table 2. Scoring Criteria for 10 Vegetation Metrics for Large

Depressional Wetlands .................................................................43

Table 3: Proposed metrics, proposed metric calculations,

and score calculations used for developing wetland

macroinvertebrate indices............................................................56
vii

Foreword

In 1999, the U.S. Environmental Protection Agency (EPA) began work on this series of reports entitled

Methods for Evaluating Wetland Condition. The purpose of these reports is to help States and

Tribes develop methods to evaluate (1) the overall ecological condition of wetlands using biological

assessments and (2) nutrient enrichment of wetlands, which is one of the primary stressors damaging

wetlands in many parts of the country. This information is intended to serve as a starting point for States

and Tribes to eventually establish biological and nutrient water quality criteria specifically refined for

wetland waterbodies.

This purpose was to be accomplished by providing a series of “state of the science” modules concerning

wetland bioassessment as well as the nutrient enrichment of wetlands. The individual module format

was used instead of one large publication to facilitate the addition of other reports as wetland science

progresses and wetlands are further incorporated into water quality programs. Also, this modular

approach allows EPA to revise reports without having to reprint them all. A list of the inaugural set of 20

modules can be found at the end of this section.

This series of reports is the product of a collaborative effort between EPA’s Health and Ecological

Criteria Division of the Office of Science and Technology (OST) and the Wetlands Division of the

Office of Wetlands, Oceans and Watersheds (OWOW). The reports were initiated with the support

and oversight of Thomas J. Danielson (OWOW), Amanda K. Parker and Susan K. Jackson (OST),

and seen to completion by Douglas G. Hoskins (OWOW) and Ifeyinwa F. Davis (OST). EPA relied

heavily on the input, recommendations, and energy of three panels of experts, which unfortunately have

too many members to list individually:

 Biological Assessment of Wetlands Workgroup

 New England Biological Assessment of Wetlands Workgroup

 Wetlands Nutrient Criteria Workgroup

More information about biological and nutrient criteria is available at the following EPA website:

http://www.epa.gov/ost/standards

More information about wetland biological assessments is available at the following EPA website:

http://www.epa.gov/owow/wetlands/bawwg
ix

List of “Methods for Evaluating Wetland

Condition” Modules

1 ................. Introduction to Wetland Biological Assessment

2 ................. Introduction to Wetland Nutrient Assessment

3 ................. The State of Wetland Science

4 ................. Study Design for Monitoring Wetlands

5 ................. Administrative Framework for the Implementation of a

Wetland Bioassessment Program

6 ................. Developing Metrics and Indexes of Biological Integrity

7 ................. Wetlands Classification

8 ................. Volunteers and Wetland Biomonitoring

9 ................. Developing an Invertebrate Index of Biological

Integrity for Wetlands

10............... Using Vegetation to Assess Environmental Conditions

in Wetlands

11 ............... Using Algae to Assess Environmental Conditions in

Wetlands

12 ............... Using amphibians in Bioassessments of Wetlands

13 ............... Biological Assessment Methods for Birds

14 ............... Wetland Bioassessment Case Studies

15 ............... Bioassessment Methods for Fish

16 ............... Vegetation-Based Indicators of Wetland Nutrient

Enrichment

17 ............... Land-Use Characterization for Nutrient and Sediment

Risk Assessment

18............... Biogeochemical Indicators

19 ............... Nutrient Load Estimation

20 .............. Sustainable Nutrient Loading

Module # Module Title
1

Florida: Monitoring Across a Nutrient

Gradient in the Everglades

Contact Russel Frydenborg

Organization Florida Department of Environmental Protection

2600 Blair Stone Road, MS 6511

Tallahassee, FL 32399-2400

Phone (850) 921-9821

E-mail russel.frydenborg@dep.state.fl.us

Purpose

T his project was initiated to monitor biological assemblages across a nutrient gradient in

the Florida Everglades in support of regulatory efforts to define a numeric water quality criterion for

phosphorus. The goal is protection of natural populations of aquatic flora and fauna in the Everglades

Protection Area.

Wetland Type

 Freshwater marshes

Assemblages

 Algae

 Macroinvertebrates

 Vascular plants

Status

 Revising sampling methods and analyzing data

Project Description

The historic Florida Everglades consisted of approximately 4 million acres of shallow sawgrass

marsh, with wet prairies and aquatic sloughs interspersed with tree islands. Today, only 50% of the

original Everglades ecosystem remains, primarily as

a result of drainage and conversion of large portions of the northern and eastern Everglades to agricultural or urban land use. The remaining portions

of the historic Everglades are located in the Water

Conservation Areas (WCAs) and Everglades National Park (ENP) (see Figure 1).

Figure 1: Area of interest within

Everglades protection area.

The Everglades ecosystem evolved under extremely low phosphorus concentrations and is considered an oligotrophic ecosystem. A large body

of evidence indicates that phosphorus is the primary limiting nutrient throughout the remaining

Everglades. Introduction of excess phosphorus to

the Everglades has resulted in ecological changes

over large areas of the marsh. The Everglades For-
2

ever Act (EFA; Section 373.4592, Florida Statutes), passed by the Florida Legislature in 1994,

stated that waters flowing into the part of the remnant Everglades known as the Everglades Protection Area (defined as Water Conservation Areas 1,

2A, 2B, 3A, 3B, and ENP) contain excessive levels of phosphorus and that a reduction in levels of

phosphorus will benefit the ecology of the Everglades Protection Area. The EFA requires the

Florida Department of Environmental Protection

(FDEP) and the South Florida Water Management

District (SFWMD) to complete research necessary

to establish a numeric phosphorus criterion for the

Everglades Protection Area.

The SFWMD Everglades System Research Division (ESRD) initiated a succession of studies, beginning in 1993 and continuing to the present, as

part of the research and monitoring being conducted

in the Everglades for phosphorus criterion development. Biological monitoring for the ESRD studies was initiated in early 1994 in WCA 2A. Data

from this and other studies are being used by FDEP

in the development of a numeric phosphorus criterion for the Everglades Protection Area.

Study Design

SFWMD ESRD initially selected 13 sites along 2

transects located downstream of canals discharging into WCA 2A and extending down the phosphorus gradient into the least affected areas of the

marsh. Sampling sites ranged from the canal inflows (discharge structures on the northeastern

margin of WCA 2A) to a site nearly 15 km downstream from the canal inflows. Three of the 13 main

sites (sites U1-U3) were specifically chosen to represent the least affected area of WCA 2A with respect to anthropogenic disturbance. A series of 15

additional “intermediate” sites were added to the

study later to obtain better spatial coverage of the

lower portion of the transects. The sites have been

monitored for water, sediment, and biological quality. Figure 2 shows the WCA 2A monitoring sites

located along the phosphorus gradient.

Sampling Methods: Algae

 Water Bottles: Phytoplankton samples initially

were collected monthly and later collected

quarterly using water bottles. Samples were

preserved in the field and sent to the FDEP

Central Biology Laboratory for taxonomic identification.

 Diatometers: Racks each containing six glass

diatometer slides were deployed quarterly at

each site. It was determined that an 8-week

deployment was necessary for sufficient periphyton growth. Diatometers were collected

and preserved and sent to the FDEP Central

Biology Laboratory for processing and taxonomic identification.

 Natural Substrate (benthic): Samples of

benthic periphyton were collected from surficial

sediment cores at the main transect sites on several occasions. Samples were retained by

SFWMD ESRD for processing and taxonomic

identification.

Analytical Methods: Algae

 Water Bottles: Samples were processed and

enumerated by FDEP Central Biology LaboFigure 2: WCA 2A monitoring

sites along the phosphorus

gradient.
3

ratory staff according to FDEP standard operating procedures (SOPs) (e.g., AB-04 and AB05; available at http://www8.myflorida.com/

labs/sop/index.htm). Analysis from this study

and other studies has indicated that Everglades

phytoplankton are largely periphyton that has

sloughed off into the water column. Thus, algal

data analysis was focused on the periphyton

data.

 Diatometers: Samples were processed and

enumerated by FDEP Central Biology Laboratory staff according to FDEP SOPs (e.g.,

AB-02, AB-02.1, AB-02.2, and AB-03; available at http://www8.myflorida.com/labs/sop/

index.htm).

 Natural Substrate (benthic): SFWMD processed and enumerated natural substrate

samples.

Sampling Methods:

Macroinvertebrates

 Dipnet: SFWMD staff conducted quarterly

macroinvertebrate sampling using a standard Dframe dipnet with a 30-mesh bag from September 1994 through November 1995. The

sampling method consisted of the collection of

20 0.5 m (in length) discrete dipnet sweeps from

representative habitats in the area of each site

on a given sampling date. The 20 dipnet sweeps

for a given site were combined and sent to the

FDEP Central Biology Laboratory for processing and taxonomic identification.

 Quan Net: Beginning in May 1996, SFWMD

staff conducted quarterly macroinvertebrate

sampling using the Quan Net method. The sampling method consisted of the deployment of a

1-m2

 frame at the site and the collection of net

samples and all vegetation within the area of

the frame. Frames were deployed in each of

several representative habitats, where present,

in the vicinity of each site. Samples from each

site/habitat were kept separate. Representative habitats were labeled as cattail, sawgrass,

or slough, depending on the predominant vegetation type. The collected material from each

site/habitat was subsampled, preserved, and

sent to the FDEP Central Biology Laboratory

for processing and taxonomic identification.

 Hester-Dendy: SFWMD staff deployed

Hester-Dendy artificial substrate samplers at

each of the main transect sites on a quarterly

basis. The samplers were deployed for a 1-

month period, after which they were collected,

preserved, and sent to the FDEP Central Biology Laboratory for processing and taxonomic

identification.

Analytical Methods:

Macroinvertebrates

 Dipnet and Quan Net: FDEP Central Biology Laboratory staff subsampled the dipnet

and Quan Net samples from each site and analyzed them according to FDEP SOPs

(e.g., IZ-02 and IZ-06; available at

http://www8.myflorida.com/labs/sop/

index.htm).

 Hester Dendy: FDEP Central Biology Laboratory staff processed and analyzed the HesterDendy samples from each site according to

FDEP SOPs (e.g., IZ-03 and IZ-06; available

at http://www8.myflorida.com/labs/sop/

index.htm).

Sampling Methods: Macrophytes

 Macrophyte Stem Density and Frequency: In

April 1997, SFWMD staff conducted a study

of macrophytes at the WCA 2A transect sites.

A 50-m tape was laid out at each transect site.

A 1-m square frame was used every 2 m along

the tape to delineate the sample area for calculation of macrophyte stem densities (stems/m2

)

and frequencies (# plots where a species was

found/total # of plots) by species.
4

 Macrophyte Harvesting: On the other side

of the 50-m tape used for establishing stem densities and frequencies, SFWMD staff harvested

macrophytes for biomass measurements, using

the 1-m square frame at five predetermined locations to mark the sample area for harvesting.

Analytical Methods:

Macrophytes

 Macrophyte Stem Density and Frequency:

Stem densities (stems/m2) and frequencies (#

plots where a species was found/total # of plots)

by species were counted at each site.

 Macrophyte Harvesting: SFWMD staff conducted biomass analysis of the harvested macrophytes for comparison of the relative biomass of several species present at each of the

WCA 2A transect sites (e.g., Eleocharis,

Nymphaea, Typha).

Lessons Learned

Periphyton, macroinvertebrate, and macrophyte

communities in WCA 2A change substantially from

reference conditions at approximately 7 to 8 km

downstream of canal discharges into WCA 2A (see

Figures 3–5). Data analysis has shown that biological populations at the two stations (E5 and F5)

nearest to the three initial reference sites (U1-U3)

are very similar in terms of biological community

structure. This suggests that these areas, despite

slight phosphorus enrichment, still support reference

condition biota. The somewhat higher phosphorus

regimes at the next stations (E4 and F4 and beyond) are associated with greater biological changes.

Experimental field dosing studies (microcosms) have

been conducted by SFWMD ESRD and show that

the addition of phosphorus causes changes in periphyton assemblages consistent with those observed

in the transect study.

The WCA 2A transect periphyton data for each

site/date have been analyzed using the entire taxonomic assemblage encountered and using lists of

pollution-sensitive and -tolerant species based on

available literature and experimental phosphorus

addition studies (the microcosms) in WCA 2A.

Macroinvertebrate data have been analyzed using

the Florida Index and the macroinvertebrate component of the Lake Condition Index (LCI), measures of the numbers of pollution-sensitive taxa in a

sample that are routinely used by FDEP in

bioassessments of streams and lakes. The use of

these methods with the WCA 2A transect data has

demonstrated a clear signal of biological disturbance

along the nutrient gradient in WCA 2A. FDEP is

using this information as well as information from

other studies conducted in the Florida Everglades

to develop a numeric phosphorus criterion for the

Everglades Protection Area.

Additional Comments

The information provided here is based solely on

the transect study by SFWMD ESRD in WCA 2A.

Research and monitoring of Florida Everglades

water, sediment and biological quality is being conducted by several research groups in WCA 2A,

WCA 1 (Arthur R. Marshall Loxahatchee National

Wildlife Refuge), Everglades National Park (ENP),

and WCA 3A, including studies by SFWMD ESRD

similar to the WCA 2A transect study.
5

Figure 3: Change point analyses of Eleocharis frequency of

occurrence and biomass data along the SFWMD transects.

Collected April 1997.

Figure 4: Results of change point analyses performed on

median total percentage of pollution-sensitive

(literature determined) periphyton taxa.
6

Figure 5: Results of change point analyses on

median Florida index values.
7

Florida: Development of a Biological

Approach for Assessing Florida

Wetland Integrity

Contact Mark Brown

Organization University of Florida, Center for Wetlands

Phelps Lab – PO Box 116350

Gainesville, FL 32611-6350

Phone (352) 392-2309

E-mail mtb@ufl.edu

Website http://www.enveng.ufl.edu/homepp/brown/syseco

Purpose

To develop “bioindicators” of ecosystemhealth for wetlands in Florida. To achieve this

goal, the project team has developed wetland

ecoregions using GIS technology, a classification

scheme for Florida wetlands, sampling protocols

for herbaceous and forested wetlands, and a quantitative index of the human disturbance gradient.

Wetland Type

 Freshwater marshes

 Forested wetlands

Assemblages

 Algae

 Macroinvertebrates

 Vascular plants

Status

Developed field protocols, sampled more than 150

wetlands statewide, developed candidate plant,

macroinvertebrate, and algae metrics for marshes.

Currently sampling and developing metrics for forested wetlands.

Project Description

The University of Florida Center for Wetlands is

involved in a multiyear wetland research project

funded by the Florida Department of Environmental Protection (FDEP) to develop an integrated biological approach for evaluating Florida’s wetlands.

The project goal is to develop an assessment approach that recognizes the utility of both biological

and functional assessments, and is rapid, reproducible, and meaningful.

The Center for Wetlands began development of

the assessment approach in 1997. Now in its fifth

year, the project, titled “Development of a Biologi-
8

cal Approach for Assessing Wetland Function and

Integrity,” has four main tasks:

Task 1. Review and synthesize all current and

relevant literature.

Task 2. Develop a wetland classification system

for wetland types in the State of Florida.

Task 3. Develop a GIS-driven methodology for

classifying bioregions within the State of Florida that

identify climatic, geologic, and geophysical provinces that are sensitive to wetland classes.

Task 4. Develop a bioassessment methodology,

biocriteria, and metrics for wetlands in the State of

Florida.

To date, Tasks 1–3 have been completed, and

work continues on Task 4. Nearly complete is a

set of metrics for herbaceous depressional wetlands,

and data are currently being analyzed to develop

metrics for forested depressional wetlands.

The development of the approach has included a

review of technical and scientific literature, a wetland classification and crosswalk, wetland

regionalization of Florida, and two wetland biological surveys in summer 1998 and summer 1999. The

biological surveys were designed to test the validity

(and necessity) of the proposed wetland regions,

to identify the appropriate biological indicator taxa,

and to quantify the gradient of human disturbance.

Study Design

The approach has included a review of technical

and scientific literature, development of a wetland

classification system and crosswalk with other classifications, wetland regionalization of Florida, and

4 years of wetland biological surveys in the growing seasons of 1998, 1999, 2000, and 2001 (a fifth

season of field surveys is under way for 2002). The

biological surveys were designed to test the validity

(and necessity) of the proposed wetland regions,

to identify the appropriate biological indicator taxa,

and to quantify the gradient of human disturbance.

The wetland classification scheme was organized

with major classes defined from three variables: (1)

dominant vegetation (forested, shrub, herbaceous);

(2) geomorphic position (stream channel [floodplain], flat topography, sloped topography, lake

fringe, depressional); and (3) primary water source

(rainfall, surface water, ground water). Subclasses

are discriminated by modifiers (hydroperiod and

plant community associations). Eleven wetland

classes were identified: forested (river swamp,

slough/strand/seepage swamp, lake swamp, depression swamp, flatland swamp); shrub dominated

(shrub-scrub swamp); and herbaceous (river marsh,

wet prairie, lake marsh, depressional marsh, seepage marsh). An HTML electronic database has been

completed that crosswalks existing wetland classifications to the new simplified classification scheme

developed for the bioassessment project.

Regionalization of the State was necessary because there is significant variation in climatic and

physical features of the Florida peninsula and it was

believed that these regional differences would equate

to variations in bioindicator “signals.” Map coverages of physical and climatic variables of the Florida

landscape were used to develop regions that had

different characteristic driving energies and landscape structural characteristics.

The map coverages were combined with GIS map

algebra to create a spatial hydrologic budget equation for the State. The equation modeled the movement of water on the landscape during the ecologically sensitive spring growing season. The output

of the model provided a value for a Potential Soil

Moisture Index (PSMI). The PSMI was separated

into four regions based on both the critical depth of

saturation and on a statistical clustering of the PSMI

values (see Figure 6). The classified regions were

tested for similarities and differences to determine if

between-region variation in wetland type and environmental variables was greater than within-region

variations.
9

The importance of hydrology in determining wetland type and location (the premise behind the

PSMI) was then tested using a hierarchical classification technique (TWINSPAN) and ordination

(DCA and CCA) with variables of seasonal and

annual rainfall, seasonal and annual potential evapotranspiration, slope, geology, drainage class, and

runoff. TWINSPAN, DCA, and CCA tested the

relationships between wetland type and climatic and

physiographic landscape characteristics. Based on

the geostatistical output, hydrology is indeed a major determinant of wetland type and location, and

supports the use of the spatial hydrologic budget

equation in delineating wetland regions.

Four years of biological surveys of wetlands

throughout the State were designed to test the

regionalization techniques as well as the metric and

bioindicator development. The 1998 pilot field research involved surveying 24 herbaceous and forested depressional wetlands in north and central

Florida. Major taxonomic assemblages were characterized and ranked along a gradient of disturbances. Sites were located within multiple land uses

such as parks, preserves, pastures, farm fields, well

fields, silviculture plots, and urban areas. Impacts

that were assessed included hydrological modifications, nutrient loading, and altered hydroperiod. The

first year of sampling resulted in development of

standardized sampling procedures, design and

implementation of a statewide sampling program,

and identification of community attributes and candidate metrics.

In the second and third field seasons, 77 herbaceous, depressional wetlands were surveyed in 3

regions (south, central, and north). Approximately

half of the wetlands were impacted (agricultural setting) and half were reference locations. Many of

the sites were paired sites (impacted and reference

at close proximity).

Figure 6: Florida’s wetland regions and field site locations.
10

In the fourth season, 72 forested, depressional

wetlands were surveyed in four regions (18 sites in

each of four regions: north, central, and southern

peninsular as well as the panhandle; see Figure 6).

Approximately ? of the wetlands were impacted (?

in agricultural settings and ? in urban settings) and ?

were reference locations. Many of the sites were

paired sites (impacted and reference at close proximity).

Quantifying gradients of human disturbance

A Land Development Intensity (LDI) index is being used to quantify gradients of human disturbance

for wetlands throughout the State. The LDI index

is calculated using land use/land cover characteristics, from aerial photographs, of lands within a 100-

m buffer surrounding the wetland. Land uses in the

area surrounding a wetland are first characterized

and then an intensity factor is assigned to each land

use type. The LDI algorithm multiplies the percent

area of each land use/land cover in the surrounding

100-m buffer by intensity coefficients. The intensity coefficients are scaled from 1 to 10 and represent intensity of environmental manipulation as measured by energy use per unit area per unit time. The

LDI index is such that lower LDI values are indicative of a lower disturbance level.

Sampling Methods: Algae

Materials

 Three 100-mL collection jars

 1-L collection jar

 Bottomless collection cup, knife, large pipette,

bag, brush/scraper

 M3 preservative

 1 L of deionized water (for dry sites)

Methods

1. Methods vary depending on whether there is

water present at the site. We are experimentally collecting dry benthic algae samples when

wetlands are dry. At this time, there is no information as to the usefulness of this sampling technique. Methods outlined below are for wet

sites.

2. Samples are taken depending on substrate and

separated as benthic, epiphyton, metaphyton,

and phytoplankton.

3. For epiphyton, the 10 aliquots are divided

equally among herbaceous and woody debris:

a. For herbaceous debris, plant stems are cut

from the soil to the water surface, and

placed in a zip-lock bag with some wetland water. The plant matter is shaken and

massaged thoroughly to dislodge the algae. Using a large pipette, 10 mL of sample

are extracted into collection jar.

b. For woody debris, a brush is used to dislodge algae. If the debris is small enough,

it is placed in a bag, similar to the herbaceous methods above. If the debris is

larger, a bottomless collection cup is used

to confine the sample, and it is brushed

while under water.

4. For benthic algae, a bottomless collection cup

is used to isolate a spot on the sediment. Then

a large pipette is used to gently stir the surface

(top 1 cm) of the sediment, and extract a 10-

mL aliquot that is placed in a collection jar.

Sampling is repeated at different locations until

a total of 100 mL is collected.

5. For metaphyton, a thumbnail size portion of the

algal mat is collected from 10 different locations throughout the wetland.

6. For phytoplankton, a total 100 mL of surface

water is collected at 10 locations throughout

the wetland.

7. All samples are preserved with M3 using 5 mL

per 100 mL of sample.

8. Samples are sent for later laboratory identification.
11

Sampling Methods:

Macroinvertebrates

Materials

 Field Physical/Chemical Characterization Data

Sheet

 Habitat Assessment Sheet

 D-Frame dipnet with no. 30 mesh

 4-L sample jars

 Buffered formalin

Methods

1. The wetland is visually examined by walking

throughout the wetland, paying close attention

to its physical and habitat characteristics, and a

Field Physical/Chemical Characterization Data

Sheet and Habitat Assessment Sheet is completed. The percent coverage of substrate type

and each habitat type is recorded.

2. A total of 20 sweeps with a D-frame dipnet are

divided between the habitat types based on their

percentage of total wetland area. Discrete 0.5-

m sweeps are performed with the dipnet. The

available substrates are sampled as determined

by the above procedures in the following

manner:

a. For areas without flow, an area of substrate that is one dipnet width wide and

approximately 0.5-m long is disturbed by

sweeping the net over the area three times

to ensure the capture of organisms.

b. For heavily vegetated areas, the net is

jabbed into the base of the vegetation, digging down to (but not into) the substrate,

and dislodging organisms using a 0.5-m

sweeping motion with the net.

c. Sand, muck, mud, and silt (nonmajor habitats) are sampled by taking 0.5-m sweeps

with the net while digging into the bottom

approximately 1 cm.

3. The number of sweeps for each habitat is recorded on the Field Physical/Chemical Characterization Data Sheet.

4. The collected samples are reduced in volume

after each discrete sample by dislodging organisms from larger debris (but retaining invertebrates in the net or sieve) and discarding the

debris. The finer debris and organisms are saved

in sample jars.

5. Samples are preserved with buffered formalin

and shipped to the FDEP Laboratory for identification.

Sampling Methods: Plants

Materials

 Field data sheets

 Compass

 100 m of tape

 FDEP’s Florida Wetland Plant Identification

Manual (Tobe et al. 1998)

Methods

1. Using a compass (or the GPS unit) four linetransects are located from four cardinal point

directions (N, S, E, W) that run parallel to the

slope of the wetland, beginning at the upland

edge (0 m) and extending into the center of the

wetland.

2. At the beginning of each transect, the approximate wetland/upland edge is located using a

combination of wetland plants and hydric soils.

3. Preferably, all four transects are set at one time,

with N, S and E, W transects meeting perpendicularly in the center, and dividing wetland into

four equal sections. Each transect is started

with the 0-m point at the wetland–upland edge

and increasing in distance toward the wetland

interior. A minimum of four data sheets is needed

per site.
12

4. Two team members walk along each transect

and record species presence of all plants within

0.5 m on either side of each 5-m interval (sampling area = 5 m2

).

5. Plant species names are recorded on the data

sheets with full genus and species if known.

6. If unknown, the species is given a unique code

identifying transect location and number of unknown encountered (N-1, N-2, N-3, E-1, E2, etc.). Voucher specimens for all unknown

species are collected, making sure to include

plant inflorescence and roots. Each specimen

is tagged with properly labeled masking tape,

and put in collection bag then pressed for later

laboratory identification.

7. Plant nomenclature follows FDEP’s Florida

Wetland Plant Identification Manual (Tobe et

al. 1998).

Sampling Methods:

Water Quality and Soils

Water quality samples are collected, preserved,

and immediately sent to the chemistry laboratory

for analysis. A composite soil sample is taken from

each vegetation zone and later analyzed for nutrients, organic matter content, and physical properties.

Analytical Methods: Plants

Field data are entered into an MS Access database for analysis. After entry, each data sheet is

checked by a second technician.

Plate 1: Cypress wetland. Photo: Elizabeth Spurrier
13

Plate 2: Macroinvertebrate sampling. Photo: Kelly Chinners Reiss
14

Maine: Developing a Statewide Biological

Monitoring and Assessment Program

for Freshwater Wetlands

Contact Jeanne DiFranco

Organization Maine Department of Environmental Protection

312 Canco Road

Portland, ME 04103

Phone (207) 822-6424

E-mail Jeanne.L.Difranco@state.me.us

Website http://www.state.me.us/dep/blwq/monitoring.htm#programs

Purpose

 Examine regional differences in wetland

macroinvertebrate and algal assemblages

 Test and refine candidate biological metrics

 Diagnose stressors and identify risks to wetlands from human activities

 Develop impairment thresholds and biocriteria

for use in State water quality standards

Wetland Type

 Freshwater marshes

Assemblages

 Macroinvertebrates

 Algae

Status

Expanding to Statewide program on a 5-year rotating basin schedule, testing and refining metrics,

analyzing data, and developing impairment thresholds.

Project Description

In 1998, the Maine Department of Environmental

Protection (DEP) began development of a biological monitoring and assessment program for freshwater wetlands. Between 1998 and 2000, DEP

conducted a pilot study in the Casco Bay watershed to develop wetland biomonitoring protocols

and identify candidate metrics related to wetland

condition. During 2001 and 2002, DEP expanded

monitoring to the Saco, Piscataqua, and Kennebec

River watersheds using the methods developed in

the pilot study. DEP uses aquatic macro-invertebrates as the primary taxonomic assemblage for this

program. Algae and diatoms are also collected as

part of a collaborative pilot project undertaken by

Dr. R. Jan Stevenson of Michigan State University

to develop algal indicators of wetland integrity.

Based on the results of Dr. Stevenson’s work, Maine

DEP will evaluate the usefulness of algae to its wetland biomonitoring program. Assessment of algae

has not been formally integrated into the Maine program at this time, however.
15

Study Design

The Maine wetland biomonitoring initiative has

been incorporated into DEP’s existing Biological

Monitoring Program, and will be extended to assess additional major watersheds Statewide. Wetland monitoring is currently coordinated with the

State’s river and stream program using the following 5-year rotating basin schedule:

Kennebec, midcoast watersheds 2002

Androscoggin watershed 2003

St. John, Presumpscot watersheds 2004

Saco, southern coastal watersheds 2005

Penobscot, downeast watersheds 2006

Considerations for site selection include hydrologic regime, geographic distribution of sites, landscape position, human disturbance gradient, management significance, and accessibility. All wetlands

sampled are semipermanently or permanently inundated, and range from minimally disturbed potential reference sites to poor-quality wetlands. As of

2002, DEP has conducted wetland biomonitoring

at 88 different sites encompassing 115 sampling

events. Some sites have been sampled repeatedly

over multiple years.

Sampling Methods:

Macroinvertebrates

Macroinvertebrates are currently sampled during

June and early July. Three different approaches have

been tested to develop both qualitative and quantitative methods. In addition, water samples are analyzed for a suite of physical and chemical parameters to help in wetland characterization, and to identify potential sources of human impact. Habitat information, dominant plant species, and a scoring of

human disturbances are recorded in the field, along

with measurements of water temperature, pH, dissolved oxygen, and conductivity.

Multihabitat sampling

A qualitative, multihabitat sampling approach was

tested, with the goal of developing a screening level

assessment tool. A standard D-frame net was used

to sample all inundated microhabitats at each site,

including emergent vegetation, aquatic macrophyte

beds, pools, and channels. Samples were “picked”

or sorted from detritus in the field. One to several

organisms representing each different taxon found

were placed into a vial of alcohol until no different

taxa were observed.

Stovepipe sampler

Maine DEP designed its own stovepipe sampler

for quantitative samples using a 5-gallon bucket with

the bottom removed. In this method, the sampler

was used to enclose three replicate plots to restrict

the movement of organisms. The stovepipe sampler was pressed into the wetland substrate, and

the contents of the sampler were then agitated.

Vegetation and surface sediment were placed into

a sieve bucket. The sampler was then swept 10

times with a small hand net. Large pieces of vegetation were washed and discarded; however, finer

plant material and detritus were retained. Samples

were preserved for later sorting and taxonomic

analysis in the laboratory.

Dipnet measured sweep

A standard D-frame net is currently used to obtain a semiquantitative sample. A sample is collected by submersing the net and sweeping through

the water column for a distance of 1 meter. The net

is bumped against the bottom substrate three times

(at the beginning, middle, and end of the sweep) to

dislodge and collect organisms from the sediment.

All material collected is placed in a sieve bucket.

Large pieces of vegetation are washed and discarded; however, finer plant material and detritus

are retained. Three replicate samples are collected

in areas of emergent vegetation. Samples are preserved for later sorting and taxonomic analysis in

the laboratory.
16

Analytical Methods:

Macroinvertebrates

Analyses performed to date reveal significant relationships between a number of candidate invertebrate metrics and watershed development. Many

invertebrate metrics tested also appear to respond

to changes in water quality typically associated with

urban nonpoint source pollution, including elevated

conductivity and concentrations of anions, cations,

and nutrients. As new data are collected, candidate metrics will be tested and refined, and regional

differences and ecological linkages among wetlands

and other waters will be examined. DEP is also

developing impairment thresholds for wetlands.

This is a necessary first step to enable the State to

use biological monitoring data in wetland management decisions and development of wetlandspecific water quality standards (designated uses

and biological criteria), and to assess wetland condition and attainment status.

Sampling Methods: Algae

Quantitative and qualitative algae samples were

gathered from the same wetland sites as used for

macroinvertebrate sampling. Four algae sample

types were tested to determine which produced the

best indicators. Samples were collected from the

water column, plant stems, and sediments. Samples

from multiple sites within each wetland were

composited into one sample from each habitat. In

addition, a multihabitat sample was collected from

each site. Samples were examined microscopically

to determine species numbers and relative abundances of different species in samples. Chlorophyll

a was quantified from a separate water column

sample as an indicator of algal biomass.

For sampling, garden shears were used to clip

plant stems below the water line. A turkey baster

was used to collect qualitative sediment samples;

however, this method was revised in 2002 to obtain a more quantitative sample. Sediment is currently collected over a known surface area using a

petri dish pressed into the substrate and retrieved

with a spatula. Three replicates are collected and

composited into a single sediment sample. A long

handled dipper is used to collect water samples.

For the multihabitat sample, a dose from each singlehabitat sample was combined into one container.

Plate 3: Maine’s stovepipe

sampler and sieve bucket.

Photo: Maine DEP

Plate 4: Picking

macroinvertebrates from

multihabitat sample.

Photo: Maine DEP
17

Analytical Methods: Algae

Dr. Jan Stevenson of Michigan State University is

using three disturbance indicators: the land use indicator developed by Maine DEP, trophic status

indicators (total nitrogen, total phosphorus, and

chlorophyll a), and hydrologic and sewage chemicals (Ca, Na, Cl). Dr. Stevenson is comparing a

suite of algal indicators to determine which types of

indicators respond to the three disturbance indicators. The algae indicators include biological integrity measures such as genus-species richness,

Shannon diversity, and number of taxa in genera.

Dr. Stevenson is also using European algal

autoecology information to determine environmental characteristics for the taxa. This information will

give an autoecological index that shows a relationship to variables such as moisture, organic N, low

oxygen, pH, salt, and nutrients. Although algae was

collected from all sites sampled between 1998 and

2002, only samples from 1998 and 1999 have been

processed to date because of funding limitations.

Lessons Learned

 Incorporating a wetland monitoring component

into Maine’s existing biomonitoring program has

been an efficient way to pool limited resources

and build on Maine’s successful river and stream

biomonitoring experience.

 To implement a comprehensive biomonitoring

program for wetlands, Maine DEP needs to

build the capacity to assess multiple biological

assemblages, including algae and vascular

plants. This capability will improve the State’s

ability to evaluate wetland impacts from stressors such as nutrient enrichment and hydrologic

changes, and will allow for the assessment of

less frequently inundated wetland types where

aquatic invertebrates are not naturally abundant.

Current funding and staff levels prohibit expansion of the program at this time.
18

Maryland: Developing an IBI Assessment for

Restored Wetlands in the Mid-Atlantic States

Contact Peter Lowe

Organization USGS BRD

Patuxent Wildlife Research Center

11510 American Holly Drive

Laurel, MD 20708-4017

Phone (301) 497-5705

E-mail peter_lowe@usgs.gov

Website http://www.pwrc.usgs.gov/wli/

Purpose

 Develop sampling methods for different assemblages

 Develop a yardstick of biological metrics to

assess the progress and condition of reconstructed wetlands in Maryland, Delaware, and

Virginia

 Compare the suitability of different assemblages

(plants, macroinvertebrates, amphibians) for

assessing wetland condition

 Evaluate the sources and magnitude of variance

in data collected for biological metrics

 Evaluate seasonal and annual biological fluctuations for the wetland sites

Wetland Type

 Delmarva Bays (depressional wetlands with

emergent, scrub/shrub, or forested vegetation)

Assemblages

 Amphibians

 Macroinvertebrates

 Vascular plants

Project History

The project is a joint effort among the USGS,

Patuxent Wildlife Research Center, the USDA Natural Resources Conservation Service Wetland Science Institute, and EPA. Project development

started in 1995 in response to a lack of information

about the success of wetland mitigation projects,

especially those associated with wetland restoration on farmlands under set-aside programs. From

1996 to 1998, fieldwork focused on a set of restored and existing wetlands. In 1999, a second

set of wetlands was studied to evaluate the robustness of the metrics developed from the first set. All

fieldwork has now been completed and metric

development is under way.
19

Study Design

Because one of the objectives was to assess the

sources and magnitude of variance in the different

measurements, we intentionally oversampled our

sites compared with what would be done under typical regional or statewide assessments. For 3 years,

work was done on a single set of 30 wetlands, including 22 restored and 8 natural wetlands or

Delmarva bays. All wetlands in the database were

depressional, semipermanent, or seasonal.

Macroinvertebrates were sampled one to four times

each year, depending on hydrology; macrophytes

during spring and late summer; and amphibians almost continuously during the breeding season.

Sampling Methods and Analysis:

Macrophytes

Data on plant species composition, abundance,

and dominance were collected through the use of

line transects. Pairs of 50 m transects were placed

on opposite sides of the wetland at four different

hydrological levels as determined at “full pool” levels. The hydrological levels were buffer, less than

25 cm, 25 to 45 cm, and greater than 45 cm. In

wetlands larger than 5 acres, the number of transect

pairs was doubled. Each transect was divided into

five runs of 5 m each, alternating with 5 m of

unsampled transect. Along each run, sampling occurred at specific points (as in point intercept methods) at 1-m intervals. Thus, each run of 5 m contained five sample points. Each transect consisted

of five runs of five sampling points each. Each hydrological depth was sampled by 2 transects, bringing the total number of sample points per hydrological zone to 50. There were:

(4 hydrological zones) H (2 transects each zone)

H (5 runs per transect) H (5 points per run)

= (200 points per wetland)

This was done in spring and fall for each wetland.

At each point, the species name and number of individuals were recorded.

In addition to point data, incidental species along

the line were recorded to include the species not

intercepted at any points along the line. These species were included in species richness and attributes

derived from presence or absence data, but not in

dominance calculations.

Sampling Methods and Analysis:

Macroinvertebrates

Aquatic invertebrates were sampled at approximately 6-week intervals beginning in late May and

continuing until October and were conducted in

association with the sampling of water quality,

aquatic plants, and hydrological and wetland dimensions. Invertebrate samples were collected along

transects following compass coordinates originating from markers placed in the deepest part of every wetland before the sampling seasons began.

Transect coordinates were randomly selected for

each wetland and sampling time. The method of

compass points was adapted to each wetland’s

morphology.

Samples were collected from three depth ranges

along the transects to determine invertebrate relative abundance, diversity, and relative biomass in

each wetland. As long as water depths were

adequate, samples were collected from along the

transects at the following water depths: less than

15.0 cm, 15.1 to 45.0 cm, and greater than

45.1 cm.

Samples were collected using a modified Gerking

box sampler, which is a sheet aluminum box with a

sliding screen door (1-mm mesh) at the bottom.

The sampler has the advantage of allowing simultaneous collection of benthic, pelagic, neutonic, and

plant-associated invertebrates. The sampler was

lowered to the floor of the sampling area with the

screen door open. The vegetation in the sampler

was then cut at the mud-water interface and put

into prelabeled plastic bags. Then the screen door

was slowly closed as the sediments just in front of
20

the advancing screen were stirred into the water

column. After the screen was closed, soil materials

were sieved through the screen by shaking the box.

All invertebrates caught on the screen were then

placed in prelabeled plastic bags. The bags of vegetation and invertebrates were placed on wet ice as

soon as possible and stored in a refrigerator until

the samples could be picked. Each individual was

keyed to the lowest taxonomic level feasible, typically species or genus. Data on species composition and abundance were used to generate metrics

in a manner similar to that for plants. An approved

quality assurance/quality control process was followed, which included independent validation of

20% of our samples.

Sampling Methods and Analysis:

Amphibians

Each site was sampled for amphibian larvae once

every 4 weeks. Sampled areas consisted of the

perimeter of the open-water portion of the wetland

and lightly vegetated areas that allowed a seine to

pass. We used a 6H8-m nylon mesh (1/16") seine

to sample each wetland by wading out 3 to 5 m

away from the shoreline and then moving in towards

the shoreline in one continuous sweep. Sampling

was time-constrained to 2 hours. If new species

were caught during the last two sweeps, the sampling period was extended until no additional new

species were found. Amphibian larvae were identified using published keys and by temporarily housing tadpoles until they metamorphosed and could

be identified.

Drift fences were also used to supplement seining

data and obtain information on adults and

metamorphs. We initially considered surrounding

each wetland with a drift fence but realized the impracticality of that idea. Therefore, each wetland

was provided with 50-cm-tall and 15-m-long drift

fences. If possible, the fences were placed along

drains, travel corridors, and other likely points of

amphibian use in order to maximize capture of individuals entering and exiting the wetlands. Five-gallon plastic buckets were buried in pairs at the ends

and midpoint on the inside and outside of the fence

to capture adults entering the ponds and juveniles

leaving the ponds. Wet sponges, rocks, and vegetation were placed in the buckets to prevent desiccation and provide some cover and refugia to captured individuals. All amphibians were identified,

sexed, and returned to the inside of the fence at the

wetland from which they were captured. Juveniles

and metamorphs departing the wetlands were identified, counted, examined for malformations, and

released on the outside of the drift fence.

General Analytical Methods

A fundamental component of this study is to devise a gradient of physical factors (e.g., land use in

drainage area, management techniques, landscape

features, method of restoration) that affect wetland

health. From there, data on the frequency of occurrence and relative abundance of species, guilds, or

trophic classes will be used to develop attributes

for an index of biological integrity (IBI) for each of

the assemblages. An attribute will be considered a

valid metric if it relates either positively or negatively to the physical gradient. We will then compare the IBIs developed and determine if they are

consistent in ranking wetlands. Once acceptable

IBIs have been developed on the initial wetland

base, we will apply them to the second set of wetlands to validate the model. In addition, the variance within our sampling methodology will be assessed.

Lessons Learned

Overall

 Many of the wetlands in our bases were only a

few years old when we started and may not

have had sufficient time for ecological and anthropogenic factors to separate them along a

physical gradient.
21

 As a result, development of a reliable and ecologically meaningful gradient has been one of

the most difficult parts of this project.

 It would be very instructive to revisit these wetlands after 10 or 15 years and see how they

have changed.

Macrophytes

 There can be considerable differences between

mid- and late summer in the ability to easily

record and identify plants. This is especially true

for graminoids, which are primarily identified

by fruiting body characteristics. In addition,

many legumes, composites, and warm-season

grasses are present in late summer but not apparent in spring.

 The inclusion of incidental species added appreciably to the number of species identified in

a particular wetland. We are evaluating whether

this inclusion has an effect on the resulting

metrics.

 Deep-water areas (greater than 45 cm) have

much lower species richness than shallower

zones and do not need to be sampled at the

same level of intensity at the same site.

 Permanent transects are preferred if data collection can continue over several years. This

will allow for the annual and seasonal changes

that occur over time through shifts in hydrology.

Macroinvertebrates

 Picking, sorting, and identifying aquatic insects

was one of the most laborious aspects of the

study. Investigators wishing to use invertebrates

in their bioassessments should allocate sufficient

resources to accomplish the task.

 Invertebrate species presence, and especially

abundance, are seasonally quite variable. June

to early July before the drying of mid- to late

summer begins seems to be the best months for

finding the greatest diversity and abundance of

macroinvertebrates.

Amphibians

 The reduced species richness of amphibians,

compared to macrophytes and macroinvertebrates, may limit the number and types

of metrics that can be developed from this

assemblage.

 Adequate sampling for amphibians requires

more trips and techniques than other assemblages. This is due to their mobility, multiple life

history strategies, and variable breeding periods among species. Sampling for one life stage

only is probably not as effective as sampling for

adults and tadpoles in determining amphibian

usage of a wetland.
22

Massachusetts: Use of Multimetric Indices to

Examine Ecological Integrity of Salt Marsh

Wetlands in Cape Cod

Contact Bruce K. Carlisle

Organization Massachusetts Coastal Zone Management

251 Causeway Street

Boston, MA 02114

Phone (617) 626-1200

E-mail bruce.carlisle@state.ma.us

Website http://www.state.ma.us/czm/wastart.htm

Purpose

The primary focus of the Cape Cod SaltMarsh Assessment Project is to advance and

improve the salt marsh assessment approach developed by the Massachusetts Coastal Zone Management (MA CZM) team through the application

and review of the existing protocol in two different

assessment investigations. The first investigation,

conducted in the 1999 field season (May to October index period), examined salt marsh indicators

from six sites on the Cape Cod Bay coast; these

sites had varying types and intensities of human land

use or disturbance. The second investigation is a

long-term comparison of indicators from three pairs

of salt marshes, each pair having a marsh area with

restricted tidal hydrology and a corresponding area

with normal tidal hydrology. The intent of this work

is to document differences in indicators before and

after tidal restoration actions. Through the implementation of these two investigations additional

objectives will be realized. The collection and compilation of data on the condition of relatively undisturbed sites is of critical importance to the evaluation and determination of impaired sites. This

project will serve to expand the salt marsh reference site database. Another important aspect of

this project will be to further examine the suite of

existing metrics and attributes used for biological

comparison and to explore new metrics, based on

the project data and literature/information base. The

long-term tide restriction study will provide insight

as to the utility of this assessment approach as a

measure of determining salt marsh restoration

progress and trajectory.

Wetland Type

 Salt marshes

Assemblages

 Birds

 Fish

 Macroinvertebrates

 Vascular Plants

Status

The Cape Cod Salt Marsh Assessment Project is

approximately two-thirds complete, with the landuse investigation finished and the longer-term tide

restriction study entering its third year (or index sea-
23

son). The report for the 1999 land-use investigation is being finalized and should be available in early

fall 2002 from this site or MA CZM’s site (http://

www.state.ma.us/czm/wastart.htm). EPA grant

funds will support two additional years of field data

collection, with project completion anticipated in

March 2004. Field investigations from 2000 through

2004 are focused on the assessment of the effects

of tidal hydrology alterations and the restoration of

normal hydrology on salt marsh ecological integrity.

The study involved three salt marsh sites with tidal

hydrological restrictions. Of the project sites, one

(EOBP) received restoration action in spring 2002,

one (EMC) is predicted for fall 2002, and one

(MSLP) is still uncertain for restoration action, with

unforeseen complexities. In addition, the project

team expects to sample one additional reference

site each year to expand the regional reference

dataset.

Project Description

Massachusetts Coastal Zone Management

launched its Wetland Assessment Program with its

first effort in wetland bioassessment in 1996-1998

at the Waquoit Bay National Estuarine Research

Reserve. A subsequent project was implemented

on Massachusetts’ North Shore in 1997-1999.

The EPA Region I sponsored pilot, Cape Cod

Salt Marsh Assessment Project, began in 1999 and

will continue through 2004. The investigative team

for this project includes Bruce Carlisle, Tony Wilbur,

Jan Peter Smith, and Jay Baker from MA Coastal

Zone Management, and Anna Hicks, an independent consultant specializing in aquatic

macroinvertebrates. Partners for this project include the MassBays Program, UMass Extension,

the MA Department of Environmental Management

(DEM) and its Area of Critical Environmental Concern Program, the Waquoit Bay National Estuarine

Research Reserve, the Cape Cod Commission, and

the towns of Barnstable, Eastham, Mashpee, Orleans, and Sandwich.

The Massachusetts CZM project team will apply

standardized sampling and surveying protocol to salt

marsh study sites to gather biological, chemical, and

physical data for the two investigations referred to

earlier.

The project team will analyze and express the biological data through a series of existing metrics or

attributes, or develop new metrics or attributes as

necessary, based on the project data and literature

base. Chemical and physical data will be utilized as

supporting information sources.

The team will make recommendations for revisions, additions, or deletions to its current wetlands

assessment approach.

Study Design

In the 1999 field season, six salt marsh sites with

varying types and degrees of intensity of surrounding land use were selected. Two sites with minimal

human land use (conservation land and no tidal hydrological alteration) were chosen as reference

sites, representing the best attainable conditions in

the immediate region.

The four other salt marsh study sites have varied

land uses including residential, commercial, and

transportation. The impacts associated with these

land uses include direct stormwater outfalls, large

impervious areas, septic systems, lawn fertilizer/

chemicals, pet waste, automobile emissions/

byproducts, and direct habitat alterations.

For the 2000 field season, three salt marshes with

tidal hydrological restrictions will be studied. Measurements will be made both at the salt marsh affected by the tidal restriction (the restricted study

site) and at the salt marsh below the restrictive feature (the reference site). The reference sites or

marshes below the tidal restriction receive normal

tidal influence and inundation. In addition, the 2000

field season will also include two additional long-
24

term salt marsh reference sites. The continued development of a robust reference site database is

critical to the continuing evolution and application

of wetland bioassessment.

Sampling Methods:

Macroinvertebrates

At each site, a habitat characterization form will

be completed that summarizes the ambient salt

marsh habitat conditions at the study site. The information collected includes water quality parameters (temperature, pH, dissolved oxygen, salinity,

and color) and habitat descriptors on hydrology,

vegetation, substrate, available food sources for invertebrates, and degree of human impact.

The sampling protocol will utilize tree devices for

application in different habitats:

Intertidal zone

Plot sampling using wooden frame (18"H18")

Subtidal (permanently flooded) zone

D-net, plot sampling, and auger

 At each sampling site, three sampling stations will

be selected over a defined linear distance of the

creek channel, stations will be located at 1/3 intervals. At each of the three stations a representative

composite sample of macroinvertebrates will be collected as follows:

Intertidal bank zone at low tide: 1 plot sample

Subtidal estuarine zone at low tide: 1 plot

sample, 1 D-net sample, and 1 auger sample

Each sample will be placed in a zip lock bag labeled with site number, site name, date of sampling,

sample number, sampling method, and name of sampler. The site field sheets also record the relevant

sample numbers. Samples will be preserved in 70%

isopropenol alcohol and placed in a cooler ready

to be transported to the laboratory for sorting and

identification.

Invertebrate samples are taken once in May and

again in August.

Sampling Methods: Vegetation

At each site, the salt marsh wetland vegetation

will be surveyed according to this protocol. Six

transects will be established based on a stratified

random sampling approach. A defined linear distance of the salt marsh creek channel is established.

The evaluation area will be segmented into three

subunits along equal sections of the creek channel.

The first third of this length is subunit #1, the second third is subunit #2, and the final third is subunit

#3. In each of the subunits, two randomly selected

transects will be laid. The transect locations will be

determined by a computer random numbers

algorithm.

The transects will run roughly perpendicular from

the channel to the upland edge, and each transect

will be laid according to a consistent compass bearing. Along each transect, 1-m2

 quadrats will be

located every 60 feet, starting at the creek edge

and progressing along the entire length of the transect

until the upland edge. The last quadrat will be located in the salt marsh fringe community, well within

the wetland and not on the upland.

Using a standard data sheet, in each quadrat along

each transect, every plant occurring within that quadrat will be identified by genus and species. For

each unique species within the quadrat, the abundance of that species will be determined using standardized coverage charts. Investigators will also

define the community type that the quadrat falls in:

low marsh, high marsh, or fringe. To be as accurate as possible, coverage estimates include duff,

leaves, bare ground, and open water, collectively

designated as “other.” Coverage estimates will be

adjusted during the analysis stage to account for

the coverage of this “other” category.
25

Vegetation surveys will be conducted once at each

site during the peak growing period from mid-July

to mid-September.

Sampling Methods: Birds

Point counts will be utilized as the primary sampling method, using visual and auditory cues. At

least two expert observers, including the principal

investigator, will sit quietly from a vantage point

where all of the evaluation area can be viewed.

Using a standard data sheet, all species and individuals will be counted and recorded by the observers, as they are heard or seen demonstrating

any activity in the evaluation area or in a 100-foot

buffer area surrounding the evaluation area. Counts

will be conducted for a period of 20 minutes, separated into four 5-minute sample intervals. All individuals will be counted, with a concerted effort not

to duplicate individuals. An additional 10 minutes

will be allotted to allow observers to walk slowly

along the perimeter of the wetland in order to detect any species not tallied in the 20-minute count.

Several sites may be visited on the same day, with

census beginning at approximately 6 am and ceasing at approximately 8:30 am in order to capture

peak activity. Sites will be sampled in late August

to capture migrating shorebird usage, as saltmarsh

habitats are known to have comparatively fewer

breeding species.

Sampling Methods: Fish

The sampling strategy will be to capture the channel habitat (sub- and intertidal). As detailed above,

the evaluation area will be divided into three subunits along a defined length of creek channel. Stations will be established for each habitat as follows.

In the creek channel, fixed stations will be determined by a computer random numbers algorithm

producing a random integer between 0 and 100.

The random integer will be the distance of the starting point for the seine haul in feet from the start

point (0’) of each subunit.

Seines will be utilized to sample the creek channel. At three stations, as defined above, the seine

will be dragged through the water column along the

creek bank and substrate for a length of 5 meters.

Seines will be carefully withdrawn from the creek

and the collected fish will be carefully extracted from

the seine into a processing bucket. The fish will

then be removed by dipnets and individuals will be

identified and measured. Abundance and total biomass by species will be enumerated. All data are

recorded on a standardized field sheet.

One sample run will be conducted each month

from April to October.

Analytical Methods

Biological data collected at wetland study sites

are compared to data collected at the wetland reference sites. Multimetric data analysis techniques

are employed to examine attributes and variables

of biological data, and these metrics are combined

into a quantitative final index. A metric is a parameter or variable that represents some feature, status, or attribute of biological assemblage, chemical

state, or physical condition. In a multimetric approach, several different metrics are chosen in order to effectively capture and integrate information

from individual, population, guild, community, and

ecosystem levels and processes. Metrics are selected on the basis of literature reviews, historical

data, and professional knowledge. The quantitative output from each metric is then combined to

produce an index. An index is the aggregate of

weighted metric scores that serves to summarize

the biological condition.

Lessons Learned

Through the three pilot projects, the MA CZM

project team has been able to learn from each application and as a result has made several small,

incremental revisions to many components of the

protocols, including adjustments to sampling meth-
26

ods and shifts in the attributes and metrics examined. Each application has also generated results

that indicate decreasing biological integrity with increasing land-use stressors.

Results from 6 years of wetland assessment

experience have led the MA CZM team to conclude that this assessment approach has significant

potential for a number of management applications,

including:

 Inventory efforts such as identifying and protecting unique and important habitat

 Identification of degraded salt marsh sites and

presence of invasive (nonnative) species in order to evaluate restoration potential and/or report on wetland status (i.e., 305b reports)

 Monitoring and evaluating the effectiveness of

restoration actions

 Long-term tracking of salt marsh condition and

documenting the effects of disturbances (i.e.,

stormwater pollution, eutrophication,

hydromodification)

Many of these applications have appeal to watershed-based organizations and agencies as well as

volunteer groups. The techniques of the MA CZM

salt marsh assessment approach have been successfully taught through a comprehensive volunteer training program on Massachusetts’ North Shore. At

the time of writing, the Wetland Health Assessment

Toolbox program is entering its fourth year, guiding

and supporting volunteer groups as they monitor

salt marsh restoration sites in their region (http://

www.salemsound.org/wetlands.htm).

Results from this Cape Cod Salt Marsh Assessment Project are still being analyzed and written

up, but some initial details are available: The 1999

land-use investigation has confirmed the trend seen

in the previous two studies where increasing levels

of land use around a given salt marsh site (wetland)

correspond with decreased ecological condition, in

this case indicated by the average of the Plant Community Index and the Invertebrate Community

Index.
27

Figure 7: Relationships between wetland ecological condition and

land use. The Land Use Index (LUI) score is a measure of human

disturbance. With increasing land use types and intensities, the LUI

score declines. Similarly, the Wetland Ecological Condition

(WEC) is a measure of biological integrity. A lower WEC score

indicates increasing biological impairment.
28

Figure 8: Two sets of multimetric goals for evaluating restoration.

The restored site is on the bottom, its paired reference (directly

below/seaward tide restriction feature) is the middle graph, and a

regional reference marsh is displayed at top.
29

Plate 5: Cape Cod, MA, salt marsh plant survey:

1 m2 along transect. Photo: B.K. Carlisle

Plate 6: Cape Cod, MA, salt marsh nekton: Watch your fingers!

(blue crab, Callinectes sapidus). Photo: B.K. Carlisle
30

Massachusetts: Involving Volunteers in

Examining the Ecological Integrity of Coastal

Wetlands in Cape Cod, Massachusetts

Contact Britta Magnuson

Organization Salem Sound 2000

201-203 Washington St., Suite 9

Salem, MA 01970

Phone (508) 741-7900

E-mail Brita.Magnuson@salemsound.org

Purpose

 Training volunteers to do biological assessments

 Comparing results of volunteers to trained scientists

Wetland Type

 Salt marshes

Assemblages

 Birds

 Fish

 Macroinvertebrates

 Vascular plants

Status

Completed 3 years of sampling. Analyzing data.

Project Description

“Salem Sound 2000” and “8 Towns and the Bay,”

two regional subcommittees of the Massachusetts

Bays National Estuary Program, participated in a

salt marsh monitoring pilot project involving citizen

volunteers in conjunction with UMass Cooperative

Extension Service and Massachusetts Coastal Zone

Management (MCZM).

During the summers of 1999 and 2000, more than

40 volunteers participated in training workshops and

field data collection for a variety of parameters:

water chemistry, land use index (a habitat assessment), aquatic macroinvertebrates, birds, tidal influence, and vegetation. During 1999, professional

scientists did independent assessments, and the

volunteers conducted assessments (using the same

sampling protocols as the professionals) with the

guidance of trained staff members. Data comparisons, as well as feedback from volunteer participants, were used to modify and improve training

protocols for the 2000 field season. Field data were

collected in 2000 at the same sites, which included

Contact Jan Smith

Organization Mass Bays National Estuary

Program

251 Causeway Street, Suite 900

Boston, MA 02114

Phone (617) 626-1231

E-mail jan.smith@state.ma.us

Website http://www.state.ma.us/czm/

wastart.htm
31

salt marsh sites that were affected by tidal restrictions and sites impacted by stormwater discharges.

The project is expected to be a model in other areas of New England. A volunteer training manual is

currently being printed and should be available at

the time of this printing. Funding from a private

foundation has enabled the program to continue for

two additional years, 2001 and 2002, and it is

planned to add fish as an additional parameter for

the volunteers to measure. The sampling protocols are summarized in the MCZM project summary.

Lessons Learned

Volunteers may need ongoing field instruction in

order to ensure quality data collection. The initial

comparison between data collected by volunteers

and data collected by professionals indicated gaps,

which improved field assistance was able to reduce.

Also, asking the volunteers to review and comment

on training workshops was helpful in improving program design. Although there is high turnover in volunteers from year to year, we are finding, after 4

years into the program, that some volunteers do

come back after a year’s break, and that the skills

of the returnees definitely improve after the first year.

Offering teacher training credits improves the interest of educators in participating in the program, and

many get interesting ideas for future use in their classrooms.
32

Purpose

Subproject 1: Coastal wetlands

 Collect baseline data on water quality and adjacent land use, as well as plant, invertebrate,

and vertebrate communities from Great Lakes

coastal wetland sites experiencing a continuum

of disturbance

 Continue development of invertebrate and fishbased indices of biological integrity (IBI’s) by

plant zone, for Great Lakes coastal wetlands

 Continue testing and validation of our IBIs

Subproject 2: Inland, forested depressional

wetlands

 Collect baseline data on plant, invertebrate, and

vertebrate communities with accompanying

chemical/physical parameters from reference

and impacted forested, depressional wetlands

of southern Michigan

 Develop an IBI for forested depressional wetlands of southern Michigan based on invertebrates, plants, fish (if present), and birds

Wetland Type

 Great Lakes coastal

 Inland, forested depressional

Assemblages

 Birds

 Fish

 Macroinvertebrates

 Vascular plants

Status

Completed 3 years of sampling. Analyzing data.

Project Description

Subproject 1: Coastal wetlands

We developed a preliminary IBI for Lake Huron

based on invertebrate data collected from coastal

wetlands with funds provided by the Michigan DeContact Thomas M. Burton

Organization Michigan State University

Department of Zoology

203 Natural Science

Last Lansing, MI 48824-1115

Phone (517) 353-4475

E-mail burtont@msu.edu

Michigan: Bioassessment Procedures and

Baseline Reference Data for Great Lakes

Coastal Marshes and Inland Forested

Wetlands in Michigan

Contact Donald G. Uzarski

Organization Grand Valley State University

Annis Water Resources Institute

Lake Michigan Center

740 West Shoreline Dr.

Muskegon, MI 49441

Phone (616) 895-3989

E-mail uzarskid@gvsu.edu
33

partment of Environmental Quality, the Nature Conservancy, U.S. EPA, and the U.S. Geological Survey and published it in the journal Wetlands (Burton et al. 1999). Since publication, we have continued testing the IBI with data from additional

Lakes Huron and Michigan fringing wetland sites.

We also have continued monitoring a set of initial

sites used in IBI development as lake levels declined in order to determine whether and how well

the IBI works as lake levels decline substantially

below levels that occurred during IBI development.

We have collected extensive data on invertebrates,

plant, fish, and bird communities on Lake Huron

wetlands since the early 1990’s. Much of our work

has been in collaboration with the Great Lakes Science Laboratory (BRD-USGS), Michigan Natural

Features Inventory (MNFI), and the Ohio Biological Survey (OBS).

Our preliminary IBI for Lake Huron fringing wetlands was developed using macroinvertebrate data

collected in 1997 from six wetlands. It was tested

using data collected from 11 Lake Huron wetlands

(6 original and 5 additional) in 1998 at lake levels

substantially lower than in 1997. We continued to

test the IBI using data collected from 12 sites (7

original and 5 additional) at even lower lake levels

in 1999 and from 5 additional sites at extremely

low water levels in 2000. Even though some plant

community zones used in IBI development were

not flooded and could not be sampled in 1999 and

2000, the IBI functioned extremely well. In 2001,

we sampled similar fringing wetlands of northern

Lake Michigan along with northern Lake Huron sites.

The IBI appears to work for fringing wetlands of

northern Lake Michigan as well.

While testing and validating the Lake Huron IBI

in 1999, 2000, and 2001, we developed modifications to simplify and improve it. A detailed explanation for the modifications was presented at “Wetlands 2000” in Quebec City in August 2000, and

additional modifications to remove the “preliminary”

status were presented in Lake Placid, New York,

in June 2002. The modifications are as follows:

1. The Typha zone should be removed from the

IBI.

2. The four diversity and richness metrics should

be calculated by plant zone instead of combining data from all plant zones before calculations

are made.

3. Two new metrics should be added to the Inner

Scirpus Zone:

– Relative abundance Isopoda (%)—

Decreases with disturbance

– Relative abundance Amphipoda (%)—

Increases with disturbance

4. Use 1⁄2 person-hour count to determine number of individuals counted per replicate. Count

either 50, 100, or 150.

A manuscript noting all of the above improvements

is in preparation and will be submitted to the journal Aquatic Ecosystem Health and Management

by September 2002 as part of a special issue on

coastal wetlands.

We are optimistic that our Lake Huron IBI will

work for northern Lake Michigan fringing wetland

sites. These sites include most wetlands along the

southern shore of the Upper Peninsula of Michigan

from St. Ignace to the Wisconsin border. Most of

these wetlands appear to be comparable in plant

community composition and structure to the Lake

Huron wetlands. Preliminary testing took place in

2001 and we expect to be able to recommend the

IBI’s use for these types of wetlands.

In 2000, we began development of new fish- and

macroinvertebrate-based IBIs for the drowned river

mouth wetlands of Lake Michigan. We collected

data from eight sites representing a gradient of anthropogenic disturbance. Ten sites were sampled

in 2001 and several new sites will be added in 2002.

We expanded our work on drowned river mouth

wetlands in 2001 to Lake Superior wetlands.
34

Subproject 2: Inland, forested depressional

wetlands

We started our forested depressional wetland

project in 1999. Invertebrates were collected three

times per year from eight forested depressional

wetlands during 1999 and 2000 using dipnets, activity traps, and black lights. Additional samples

were collected from a subset of these and some

additional wetlands in 2001. Up to 40 additional

sites will be added in 2002, with emphasis on obtaining a greater array of impacted sites and extending the sites to isolated depressional wetlands

in the coastal zone of the Great Lakes. Invertebrates are identified to the lowest taxonomic unit

possible, and preliminary IBI development has begun. Accompanying chemical/physical samples

taken from surface water and mini-piezometers have

also been recorded and analyzed for potential

metrics. Comparisons of the plant and invertebrate

communities have been made using a subset of the

sites.

A list of potential metrics, a summary of our very

preliminary analyses, and conclusions that we drew

from the 2000 data set are as follows:

Significantly higher at reference sites (MannWhitney tests):

 Isopoda p < 0.001

 Amphipoda p = 0.004

 Diptera

 Culicidae p = 0.004

 Ephemeroptera p = 0.015

 Trichoptera p = 0.078

Significantly higher at impacted sites (MannWhitney tests):

 Gastropoda

Lymnaeidae p = 0.004

Planorbidae p < 0.001

 Diptera

Chironomidae p = 0.074

Chaoboridae Collected from only one

site

 Odonata

Libellulidae p = 0.039

 Hemiptera

Lleididae p < 0.001

 Coleoptera

Halipidae p < 0.001

 Annelida

Hirudinea Collected from only one

site

Summary

Of the measured chemical/physical variables, only

depth separated impacted and reference sites.

Canopy cover was not measured in 1999 and 2000

but may have also separated impacted and reference sites. Estimates of canopy cover are currently

being obtained for all sites. Eleven (+2 ?) taxa

showed potential as metrics, but not during early

inundation (April). During full inundation (May),

the correspondence analysis grouped the wetlands

using invertebrate community composition into distinct categories.

 Surface runoff-influenced impacted sites

 Groundwater-influenced reference sites

 Precipitation-influenced reference sites

Conclusions

 Increased runoff may increase water depth and

subsequently kill trees, opening the canopy.

– Some taxa such as Libellulidae may be

responding to these changes in ambient

conditions.
35

 Most traditional chemical/physical variables will

not be useful as indicators of anthropogenic disturbance in these wetlands. These variables include turbidity, DO, pH, SRP, NH4, conductivity, alkalinity, temperature.

 An invertebrate-based IBI for (hardwood) forested wetlands of Southern Michigan appears

to be feasible.

Study Design

Subprojects 1 and 2

Sites that experience a wide range of anthropogenic disturbance, or stressors, are chosen from each

hydrogeomorphic class or subclass of wetland. The

extent of disturbance is determined using surrounding land use/land cover and limnological data. Initially, correspondence analyses of invertebrate community composition were used to determine if reference sites separated from impacted sites. When

they did, individual taxa containing the most inertia

responsible for the separation were deemed potential metrics. Mann-Whitney U tests were then

used to determine if densities of these taxa at reference sites were significantly different from densities

at impacted sites.

We use medians in place of means in the IBI because medians are more resistant to the overwhelming effects of outliers. Our goal is to typify the wetland. If an area is sampled that is depleted or concentrated in the constituents of a metric, the area

may be isolated from anthropogenic disturbance,

receiving a dose of disturbance not typical of the

entire wetland or vegetation zone, or may contain

some “natural” chemical/physical component that

is unique. Regardless of the cause, the area is not

representative of the entire wetland. The influence

of these outliers can be dampened by using the

median in place of mean as a measure of central

tendency.

After potential metrics were developed, principal

components analysis (PCA) was used to establish

principal components (PCs) based on chemical/

physical parameters as well as surrounding (1-km

buffer) land use/cover data. Pearson correlations

were done between individual metrics and PCs to

establish stressor-ecological response relationships.

PCs were then decomposed to explore relative

contributions of individual stressors.

Sampling Methods

Subproject 1: Coastal Wetlands

Macroinvertebrate samples. Macroinvertebrate

samples were collected with standard D-frame,

0.5-mm mesh dipnets. All major plant community

zones were sampled at each site, including a deep

emergent and a shallow, wet meadow zone. If certain depths contained more than one dominant plant

community along the shoreline, each plant community type was sampled.

Dipnet sampling entailed sweeps through the surface and middle of the water column and above the

sediment surface to ensure that an array of microhabitats were included in each sample. Dipnets were

emptied into white enamel pans, and 150 invertebrates were collected by removing all specimens

from small areas of the pan. Special consideration

was made to ensure that smaller organisms were

not missed, as there is a bias toward larger, more

mobile individuals with this technique. Plant detritus was left in the pan and sorted through for a few

additional minutes to ensure that sessile species were

included in the sample. If 150 individuals were not

obtained after 1⁄2 person-hour of field picking, we

collected to the next multiple of 50. The timed count

was used to semiquantify sampling effort so that it

could be used as a metric. Three replicate samples

were collected in each plant zone to obtain a measure of variance.

Dipnet samples were collected from late July

through August. Samples taken from ice-out through

mid-July generally contained less diversity and a

greater proportion of early instars of aquatic insects,
36

making identification more difficult. The July–

August time period also corresponded to the time

when plant communities, characteristic of these

wetland systems, achieved maximum annual

biomass.

Fish sampling. Six fyke nets (0.5-in mesh) were

set in each wetland for 1 net-night. Nets were set

adjacent to specific plant zones with the leads bisecting the sampling area. Minnow traps were used

as a secondary method.

Subproject 2: Inland, forested depressional

wetlands

Invertebrate sampling. Macroinvertebrate

samples were collected with standard D-frame

dipnets containing a 0.5-mm mesh. Three replicate

samples were collected from three locations in each

wetland: (1) the deepest portion (usually the center), (2) near the upland, and (3) between these

two areas. We attempted to incorporate habitat

heterogeneity by sampling as many plant zones as

possible at each location.

Dipnet sampling entailed sweeps through the water column at the surface, middle of the water column, and above the sediment surface. Dipnets were

emptied into white enamel pans and 150 invertebrates were collected by focusing on small areas of

the pan and removing all specimens. If 150 individuals were not obtained after 1⁄2 person-hour of

field picking, we collected to the next multiple of

50. The timed count was used to semiquantify sampling effort so that it could potentially be used as a

metric. Invertebrate sampling was conducted during early (April), full (May), and late inundation

(June) at each site.

Fish sampling. The temporary pools that dominate most depressional forested wetlands are unlikely to contain fish. Thus, fish sampling was only

conducted for permanent pools in depressional

wetlands when they occurred. Small fish traps were

placed in each of the permanent pools.

Birds surveys. Bird communities were surveyed

using 10-minute, 50-m radius point counts (Ralph

et al. 1995) by dual observers to sample the bird

communities at each count location. The 10-minute

counting period began when the observers reached

the perimeter of the 50-m radius so that any birds

flushed or silenced by the observer’s approach

were detected (Riffell et al. 1996). The bird communities of 30 forested wetlands were sampled in

2000 using a total of 6 count visits. A variety of

habitat measures from each site were obtained as

detailed under plant sampling below, and a manuscript on results is currently being prepared. No

new sampling has occurred since 2000.

Plant sampling. The plant community and other

habitat variables were described for each 50-m radius bird count area along four habitat sampling radii radiating from the center of each point count station following procedures detailed in Riffell et al.

(1996). The understory habitat was described using a Wien’s pole (Rotenberry and Wiens 1985)

following procedures of Riffell et al. (2001), modified as appropriate to adapt them to forested

habitat.

More traditional plant sampling, of a subset of

forested wetlands, was done by Mike Kost and

Dennis Albert of the Michigan Natural Features Inventory during 1999 and 2000 using quadrat sampling along transects running through each wetland.

They have submitted a report on their results to the

Michigan Department of Environmental Quality including recommendations on potential metrics for

these wetlands.
37

Contact Mark Gernes

Organization Minnesota Pollution Control

Agency (MPCA)

Environmental Outcomes Division

520 Lafayette Road

St. Paul, MN 55155

Phone (651) 297-3363

E-mail mark.gernes@pca.state.mn.us

Website http://www.pca.stat.mn.us

Minnesota: Developing Wetland Biocriteria

Contact Judy Helgen

Organization Minnesota Pollution Control

Agency (MPCA)

Environmental Outcomes Division

520 Lafayette Road

St. Paul, MN 55155

Phone (651) 296-7240

E-mail judy.helgen@pca.state.mn.us

Website http://www.pca.state.mn.us

Purpose

 Develop the tools to assess wetland condition

by studying vegetation and invertebrates in wetlands across a range of human disturbance

 Develop two separate Indexes of Biological

Integrity (IBI) for Minnesota depressional wetlands

Wetland Type

 Depressional wetlands, large and small; riparian wetlands

Assemblages

 Macroinvertebrates

 Vascular plants

Status

Established indexes of biological integrity (IBIs) for

plants and macroinvertebrates. Report completed

on IBIs for 44 large depressional wetlands in May

2002. In 2002, evaluating data on statistical analysis of sampling methods in 9 (invertebrates) and 12

(vegetation) large depressions. Beginning June

2002, testing IBIs in new ecoregion, comparing IBI

assessments with rapid assessment method and citizen monitoring data. Training of many citizen community teams in biological assessment of local wetlands continued in 2002; a pictorial guide will be

produced in 2002.

Project Descriptions

Minnesota has been developing its wetland biological assessment program since 1992.

1992 – Small depressional reference wetland

study

The first project was funded by the State legislature to develop biological reference conditions for

small depressional wetlands in central Minnesota.

Subsequent funding is primarily from EPA. The initial research studied the quantity and quality of

macroinvertebrates primarily in highest quality, least

disturbed reference sites and relatively few disturbed

depressional wetlands.

1995 – Expanded small depressional wetland

study

A second project funded by EPA was undertaken

to develop multimetric IBIs for depressional wetlands in central Minnesota. During the 1995 sampling season, MPCA collected data on macroinvertebrates and vegetation for a larger set

of depressional wetlands than the 1992 study, rep-
38

resenting a wide range of impairment. MPCA’s

report on the 1995 IBI development was completed

in 1999 and will be available on MPCA’s web site

(www.pca.state.mn.us).

1999 – Large depressional wetland study

MPCA sampled large depressional wetlands, including duplicate sampling at six of the sites, to validate metrics developed with smaller depressional

wetlands in 1995. MPCA also developed a scoring system for human disturbance (HDS) that incorporates estimates of disturbances in the buffer

area and in the near-wetland landscape, plus ranges

of chemical pollution and alterations within the wetland. The HDS scores were used as the x-axis to

calibrate the metrics and the IBI scores. Both the

vegetation and the invertebrate IBI scores show significant relations to the human disturbance scores

and to various water and sediment chemical factors. The invertebrate IBI was significantly related

to HDS, turbidity, phosphorus, chloride, and other

factors. The vegetation IBI was significantly related to HDS, phosphorus, and chloride in water;

and to copper; zinc; and nickel in sediments. Minor changes were made in metrics compared with

those used for the smaller depressions. The final

report for this project, Indexes of Biological Integrity (IBI) for Large Depressional Wetlands in Minnesota, by M.C. Gernes and J.C. Helgen, was completed in May 2002. In 1999, 27 wetlands in

riparian areas of small and medium-sized streams

in the St. Croix River basin were sampled for the

vegetation IBI. There data are currently under

analysis.

2001 – Statistical assessment of wetland

monitoring methods

MPCA sampled nine wetlands in three locations

to test the methods for the invertebrate and vegetation IBIs with statistical procedures. These data

are currently under analysis.

2002 – Citizen training

In 2002, MPCA expanded efforts for training citizens in biological assessment of wetlands, training

90 citizens in the invertebrate and vegetation methods (see Minnesota case study entitled “Dakota

County Wetlands Health Evaluation Project”). A

final guide for biological monitoring of wetlands by

citizens will be produced by MPCA in 2002. This

guide includes pictorial keys to wetlands invertebrates and plants. MPCA also trained school teachers in the assessment of ephemeral wetlands in the

spring of 2001 and 2002.

In the summer of 2002, MPCA is validating the

IBIs for depressional wetlands in 40 wetlands in

southern Minnesota to test regional applicability of

the methods. In addition, about 10 wetlands monitored by citizens will be assessed by MPCA using

the technical IBIs. The data from the assessments

by citizens using the modified IBIs and MPCA’s

technical IBIs will be compared. The IBI results

will be compared with results from a Minnesota

rapid assessment method carried out by consultants on the same 10 wetlands.

Study Design

In the 1992 project on reference wetlands, 32

least disturbed and 3 known disturbed wetlands

were sampled for macroinvertebrates. In the second research phase in 1995, 27 wetlands were

sampled for macroinvertebrates and vegetation.

The 27 wetlands represented the full range of human disturbance typical of wetlands in this part of

Minnesota, and were located in the North Central

Hardwood Forest ecoregion (Figure 9). The sites

included 6 least impaired reference sites and 21 sites

highly impacted from human disturbances such as

stormwater (12 sites) and agricultural influences (9

sites).

In the 1999 large depressional wetlands project,

44 sites were sampled, 6 of which were sampled

twice on the same date. Included were 14 highquality reference sites, 14 agriculture-influenced

sites, and 16 urban wetlands (Figure 10). These

wetlands were selected to represent the widest range

of disturbance.
39

In the 2001 statistical assessment of wetlands

methods project, three sectors of nine wetlands

were sampled for invertebrates and vegetation.

In 2002, approximately 50 large depressional

wetlands representing a gradient of human influence

are being assessed. Approximately 30 wetlands in

the Western Cornbelt Plains and the Northern Glaciated Plains ecoregions are being analyzed to determine if the IBI developed in the North Central

Hardwood Forest (NCHF) ecoregion can be used

in other regions of Minnesota. Five of these sites

will have duplicated sampling. Ten wetlands from

previous projects in the NCHF ecoregion will be

analyzed. Another 10 wetlands in the NCHF

ecoregion are being sampled by citizens using the

IBIs and by consultants using a rapid assessment

method.

In the projects in 1995, 1999, 2001, and 2002,

both invertebrate and vegetation sampling were

done. Water chemistry samples were taken in June,

Figure 9: Locations of smaller depressional wetlands

sampled in 1995.

Figure 10: Locations of the 44

large depressional wetlands

sampled in 1999.
40

and sediments were cored for metals and other factors in later summer.

In addition to invertebrate and vegetation sampling, wetlands were sampled for water pH, conductance, turbidity, dissolved oxygen, temperature,

calcium (hardness), chloride, total suspended solids, total phosphorus, and total nitrogen. Sediments

were analyzed for 15 heavy metals using ICP methods, as well as for total organic content, textural

classes, carbonates, chloride, total phosphorus, and

total nitrogen. Low-altitude aerial photographs were

taken to support the scoring for the human disturbance gradient.

Sampling Methods: General

Considerations

Stratification of the habitat for each study site was

done so as to minimize the biological variability

among the different strata of the wetland. For the

depressional wetland projects, the following habitats were identified: nearshore emergent zone, open

water submergent zone, and floating plant zone.

Sampling Methods:

Macroinvertebrates

Sampling was done in the nearshore emergent zone

during the seasonal index period of June to early

July. This time frame ensured that optimal species

maturity and richness were present. In previous

fieldwork, MPCA had determined that sampling in

May was too early because some invertebrates are

too immature for identification. Once collected, invertebrate samples were preserved and analyzed

in the laboratory. Macroinvertebrates were sampled

using both the dipnet and the activity trap method.

Dipnetting captured the greatest richness of invertebrates and the activity trap captured the active

swimmers and night-active predators.

Dipnet

The D-frame aquatic dipnet with 600-micron mesh

net was used. Two dipnet samples were taken within

the emergent vegetation zones. A 1⁄2O wire screen

fixed to a wooden frame was used to keep the vegetation from the sample. After sweeping the net

strongly through the water column four to five times

and downwards to near the bottom, the contents of

the dipnet are emptied onto the framed wire screen.

The frame is set over a pan containing sieved water

to catch invertebrates as they drop down from the

vegetation. For approximately 10 minutes, the vegetation is gently spread and invertebrates are encouraged to drop or crawl down to the water in the

pan. After separation from the vegetation, the water is then poured through a 200-micron sieve to

concentrate the sample before preservation. Preservation of the samples is done using 80% ethanol,

final concentration. Using a squirt bottle with the

alcohol solution, the sample is back-flushed into the

sample jar and labeled for later picking and identification.

Activity trap

The bottle trap works as a passive funnel trap that

collects organisms as they swim into the funnel and

pass through the neck into the bottle. Made from a

clear 2-L round-bottomed plastic beverage bottle,

the traps are nearly invisible underwater. The traps

are supported on a 4.5N wooden dowel with a flexible half section thin wall PVC pipe that allows raising and lowering the bottle trap on the dowel.

Ten bottle traps were placed in the emergent vegetation zone and left overnight for two consecutive

nights. Placement of the bottle traps was from the

nearest shallow shore edge to the inner side of the

deeper emergent vegetation zone in water no greater

than 1 meter. In the shallowest water, the traps

were placed on the bottom just under the surface

of the water. Traps were placed horizontally about

15-20 cm under the surface. The bottle traps were

back-filled with water leaving no air bubbles inside

to reduce the amount of predation within the trap.
41

Analytical Methods:

Macroinvertebrates

After the invertebrates were sorted in the lab, the

entire sample was identified and counted. The data

were entered into an ACCESS database and

scored for 10 metrics, which represent different

measures of the invertebrate community. The metrics

used for small depressions are in the 1999 report

of the 1995 project. The metrics used for large

depressions are in the 2002 report on the 1999

project (Table 1). For the 1995 data, the metrics

were validated by plotting them against a ranking of

the site disturbance based on professional judgment

and against selected chemical variables. For 1999,

the metric data were plotted against the human disturbance gradient scores (HDS) and some chemical factors to evaluate metric responses. In addition, linear regressions were done on metric data

and IBIs against the measures of human disturbance.

Chemical (e.g., phosphorus, nitrogen, chloride,

and heavy metals) and biological data were analyzed for statistically significant relationships to the

metrics and IBIs. Of the 10 invertebrate metrics,

intolerant taxa, chironomid taxa, and total taxa

showed the strongest responses to the estimated

disturbance gradient and water chemistry factors,

followed by the Odonata and ETSD metrics. HDS

scores, turbidity, and phosphorus and chloride in

water were most significantly related to the invertebrate IBI; copper in sediments was significant.

Sampling Methods: Vegetation

Vegetation sampling techniques vary greatly for different wetland habitats and study designs. MPCA

used a releve method for sampling vegetation. The

releve method was chosen for several reasons. The

primary reason is that it is easily adapted to widely

varying habitats and vegetation community structure, which is typical for depressional wetlands. This

adaptability in sampling methodology is needed for

wetlands that receive significant quantities of water

during storm events. A second reason that MPCA

selected releve sampling over line transect or quadrat

sampling was that the Minnesota Department of

Natural Resources’ Natural Heritage Program and

County Biological Survey use releves, and

Minnesota’s academic researchers also use a similar releve method for collecting vegetation community data.

All vegetation sampling was done in July, which

represents the typical period of maturity and the

best time in Minnesota for determining community

structure. In each wetland, a 100-m2 plot was established in representative sampling locations within

the emergent and open water submergent zones.

After establishing the releve plot, the vegetation

cover classes were determined for each plant species occurring in the plot. Voucher specimens were

collected at least once during the project for each

species or taxon encountered. All taxa that couldn’t

be identified reliably to the species level in the field

were also collected.

Analytical Methods: Vegetation

Ten vegetation metrics were developed and validated, using methods similar to those for the invertebrate IBI. Each promising attribute of the plant

community was plotted against a disturbance gradient. In the 1995 small depressional wetlands

project, the disturbance gradient index was developed from professional judgment ratings of several

disturbance factors including stormwater input, agricultural practices, quality of adjacent buffers, hydrologic alterations, and historical disturbances. For

the 1999 large depressional wetlands project, the

HDS scores and chemical factors were used. Metric

scoring criteria were then developed for the strongest responding metrics (Table 2). Metrics were

also plotted against chemical variables to demonstrate their response to traditional water chemistry

concerns.
42

Table 1: Scoring Criteria for 10 Invertebrate Metrics for IBI for

Large Depressional Wetlands

The 10 vegetation metrics showed significant responses to water (chloride and phosphorus) and

sediment chemistry (zinc, copper, and nickel). Sensitive species were found to be the strongest and

most reliable vegetation metrics.

Sampling Methods:

Water and Sediment

Sampling water and sediment chemistry was conducted by MPCA staff. Water analysis was done

by the Minnesota Department of Health, and sediment analysis was done under contract with the

University of Minnesota Soils Analytical Laboratory. See April 1999 report for sampling methods

for sediment and water chemistry.

Lessons Learned

 Vegetation IBIs show great promise for future

applications in wetlands biological assessment.

The wetland plant community is biologically rich

and sensitive to a variety of human disturbances.

Data are acquired in a short time.
43

Table 2: Scoring Criteria for 10 Vegetation Metrics for Large

Depressional Wetlands

 Biological metrics respond to many stressors

and to other disturbance factors in the landscape and in physical aspects of the wetlands

that can be readily measured.

 After applying appropriate stratifications, we

find that wetlands are not chaotic or highly variable. They show clear patterns and predictable responses to human disturbances.

 The invertebrate community is sensitive to many

disturbances in wetlands. Responses differ from

stream invertebrates because wetlands invertebrates are adapted to daily cycling of dissolved

oxygen in wetlands. What may be a “pollution

tolerant” invertebrate in streams may be a specialist in the wetland habitat.
44

Minnesota: Dakota County Wetland Health

Evaluation Project

Contact Daniel Huff

Environmental Education Coordinator

Organization Dakota County Environmental Education Program

4100 220th St. West, Suite 101

Farmington, MN 55024-9539

Phone (651) 480-7734

E-mail daniel.huff@co.dakota.mn.us

Website http://www.extension.umn.edu/county/dakota/Environment wetlands/wetld.html

Purpose

 Evaluate wetland health using biological data

gathered by citizen volunteers using approved

techniques developed by the Minnesota Pollution Control Agency

 Increase biodiversity in wetlands in urban areas by installing Best Management Practices

(BMPs)

 Conduct a public wetland education effort

through seminars, workshops, field days, and

media

Wetland Type

 Depressional wetlands

Assemblages

 Macroinvertebrates

 Vascular plants

Status

Since 1997, the project has operated every summer. In 2002, the program consisted of 15 monitoring teams representing 19 communities in Dakota and Hennepin Counties.

Project Description

The Dakota County Wetland Health Evaluation

Project (WHEP) uses sampling methods and evaluation metrics developed by the Minnesota Pollution Control Agency (MPCA). The project started

in 1997 and was conducted by MPCA and Minnesota Audubon. A total of 30 wetlands were monitored by 5 citizen teams representing the Minnesota Zoo, Dakota County, and the cities of

Burnsville, Eagan, and Lakeville. MPCA staff

trained volunteers in sampling protocols, quality

assurance, and plant and macroinvertebrate identification. The sampling methods are a scaled-back

version of the Minnesota Pollution Control Agency

Wetland Bioassessment Program (see “Minnesota:

Developing Wetland Biocriteria” in this module).

Each citizen team worked under the direction of a

local teacher or nature center staff. The time commitment for volunteers was approximately between

40-50 hours per year, including training, field work,

and analysis.
45

In 1998, the project expanded to eight citizen

teams, and a technical consultant was hired to conduct MPCA’s full sampling methods to facilitate

comparisons. Results of the comparison indicate

that the volunteer assessments, although not as rigorous as the professional assessments, provide repeatable results that are consistent with the more

detailed assessments. However, the volunteers tend

to score the high-quality wetlands too low because

they are unable to identify as many organisms (e.g.,

species of Carex) resulting in lower scores for the

richness metrics. A detailed description of this

project is provided on the web site listed under the

contact information (http://www.extension.umn.edu/

county/dakota/Environment/wetlands/wetld.html).

In 1999, the funding source changed from EPA

grants to funding from the Minnesota State Legislature through the Minnesota Environment and Natural Resources Trust Fund. The Minnesota State

Legislature approved continued funding for this

project for the period of 1999 to June 2002. For

the 1999 season, the project expanded to a total of

10 cities. An additional 11 wetlands were assessed

for the first time and 24 previously sampled wetlands were resampled for trend analysis. The invertebrate and vegetation IBI scores were generally consistent for each wetland, although the invertebrate score was slightly lower on average than

the vegetation scores. Each team performed a crossheck on a wetland monitored by another team. In

addition, the technical consultant field checked 4 of

the 35 wetlands sampled by the volunteer teams.

The volunteer results were compared to MPCA

standard sampling method results and were consistent for most samples.

In 2000, 10 city teams monitored 38 wetlands.

Invertebrate scores were generally lower than vegetation scores, a more significant difference than the

previous year. Below-normal precipitation for the

previous fall, winter, and spring may account for

the poor showing of invertebrate populations. Volunteers continued to use MPCA protocols and be

trained by the MPCA scientists who developed the

protocols. Eight of the 10 teams performed a crosscheck on a wetland monitored by another team.

Of these, six of the eight sites showed consistent

scores for the invertebrate index and seven of the

eight sites showed consistent scores for the vegetation index. The technical consultant field-checked

4 of the 38 wetlands sampled by the volunteer teams.

In general, the citizen data were consistent with

consultants’ findings.

In 2001, 10 teams representing 11 cities monitored 41 wetlands. A wet spring may have contributed to higher invertebrate scores than the 2000

monitoring season. Vegetation scores were consistent with previous seasons’ scores. Twenty-eight

of 39 wetlands sampled for both vegetation and

invertebrates were considered to have consistent

scores between the 2 indexes. Seven city crosschecks were performed using the invertebrate

metrics. Of these seven, five resulted in similar

scores. Six city cross-checks were performed using the vegetation metrics. Of these six, four resulted in similar scores. The technical consultant

field-checked 3 of the 41 wetlands sampled by the

volunteer teams. The consultant’s check resulted

in identical scores for two of the three volunteer

teams and was very similar for the third for the invertebrate metrics and the vegetation metrics. This

showed higher consistency between the citizen teams

and the professional cross-check than in previous

years. In addition to the 10 Dakota County teams,

3 teams within Hennepin County monitored 14

wetlands with funding from U.S. EPA and the

Minnehaha Creek Watershed District.

In addition to wetland monitoring, a wetland

remediation project was begun in 2001 at one of

the wetlands previously monitored by the project.

Cedar Pond in Eagan, MN, had scored among the

lowest of all county wetlands for both vegetation

and invertebrates when sampled by volunteers in

2000. In cooperation with the City of Eagan, a

retaining wall surrounding the pond was removed,

the slope was regarded, and three zones of native

wetland vegetation, emergent, wet meadow, and
46

upland buffer, were planted around the wetland.

Three rainwater gardens were constructed and

planted to receive and filter runoff from an adjacent

city street. WHEP volunteers participated in planting the rainwater gardens and trees along the upland buffer.

Complete reports of the 1999, 2000, and 2001

monitoring season results can be found on the web

site given above. Information on the 2001 monitoring season within Hennepin County can be found

at http://www.hcd.hennepin.mn.us/whep.html.

In 2002, communities sponsoring teams are providing funding for the project. Nine teams representing 11 cities in Dakota County and 6 teams representing 4 cities and 1 watershed district in

Hennepin County are participating in the Project.

Results from the 2002 monitoring season for both

counties will be available on the web in February

2003. The project is expected to continue.
47

Minnesota: University of Minnesota’s

IBI Development

Contact Sue Galatowitsch

Organization University of Minnesota

Department of Horticultural Science

305 Alderman Hall, 1970 Folwell Ave.

St. Paul, MN 55108

Phone (612) 624-3242

E-mail galat001@maroon.tc.umn.edu

Website http://www.hort.agri.umn.edu/mnwet/begin.htm

Purpose

Develop assessment methods to evaluate ecological condition of wetlands

Wetland Type

 Variety

Assemblages

 Amphibians

 Birds

 Fish

 Macroinvertebrates

 Vascular plants

Status

 Analyzing data and writing reports

Project Description

Developing indices of biological integrity (IBIs)

for Minnesota was pursued by researchers at the

University of Minnesota to enable quality assessments of existing and restored wetlands. Eight

series of 15 wetlands (120 sites) were used to develop wetland IBIs. Each series covers a major

wetland type in the State and is composed of reference sites (unaltered wetlands in an unimpaired setting), sites surrounded by land use typical of the

region, and sites that are highly altered. Plants, birds,

fish, invertebrates, and amphibians were surveyed

to select the best IBIs for each series.

To identify possible patterns in biological communities that may relate to land use differences, each

organismal data set (except amphibians-few organisms encountered during surveys) was explored with

TWINSPAN. TWINSPAN organizes data so that

the most similar sites (as described by their species) are grouped together as columns on a table,

and so that the species with similar habitat affinities

(as described by sites where they occur) are grouped

together as rows. This TWINSPAN table was used

to generate a list of potential indicators for analysis

with land-use data. Species or groups of species

that appeared preferential to sites with similar land

use characteristics were deemed to be potential indicators. Other common ecological measures, such

as richness (number of species), were routinely included in the list of potential indicators, as well.

Twenty-eight potential indicators identified for this

series are listed below. Proportional indicators for

animals are calculated as a total of all organisms
48

observed (not a proportion of taxa) unless noted.

Absolute abundances for plant species (vegetation)

cannot be reliably estimated from cover class data.

Importance values, an approximate measure of

abundance, were calculated by summing cover class

scores (r=0.1, +=0.5, and classes 1-5).

Potential indicators

Amphibians: None

Birds (12): Species richness (BSR); total abundance of all birds (BABU); number of wetland taxa

(BWR); proportion of wetland birds (PWB); abundance of brown-headed cowbirds (ABHC); proportion of forest-nesting birds (PFB); number of

taxa with large territories (BLTR); proportion of

insectivorous birds (PBIN); abundance of marsh

and sedge wrens (CIS); abundance of yellow

throats, swamp sparrows, and LeConte’s sparrows

(SSYT); number of open ground nesting species

(BOGR); proportion of open ground nesting species (PBOG).

Fish: None

Invertebrates (4): Total abundance (IABU), taxa

richness (ISRI), number of snail species (GASR),

proportion of snails (PGAS).

Vegetation (12): Species richness (VSR), invasive perennial species importance (IPI), number of

Carex species (CAR), importance of Carex species (CAI), number of native herbaceous perennials (HPNR), importance of native herbaceous perennials (HPNI), number of native perennial

graminoids (GPNR), importance of native perennial graminoids (GPNI), number of introduced species (INR), importance of introduced species (INI),

ratio of graminoids to herbaceous species (VGH),

ratio of annuals to perennials (VAP).

Relationship of potential indicators to landuse measures

Values for each potential indicator (PI) were calculated for each site in the series. PI values were

correlated with land use data: site alteration score,

land use cover at 500 m, 1,000 m, 2,500 m, and

5,000 m radii. For the radii data, six correlations

were calculated, one for each land cover category

(agriculture, urban, disturbed, forest, grassland, and

wetland). For this series a total of 700 relationships were tested. Relationships with Pearson correlation coefficients greater than 0.53 (p < 0.1) are

worthy of further consideration as indicators of

wetland quality. Each of these relationships were

plotted to detect if the high coefficients were based

on outliers. Those with outliers were not considered significant.

Eighteen of the original 28 potential indicators

(64%) were found to have a high correlation to land

use. Seven of the bird PIs show a land-use relationship. Birds with large territories (BLTR) are

more common to sites with less agriculture and disturbance at the regional scale (2,500-5,000 m).

Likewise, regional patterns of urbanization are negatively associated with wetland bird richness (2,500

m). Wetland bird richness is greater on sites with

more wetlands at this same scale. Overall bird richness is lower with more surrounding urbanization at

most scales. In contrast, most of the seven promising vegetation PIs show stronger land-use relationships at local scales. Introduced species (INI, INR)

and annuals (VAP) are more common on sites that

are immediately impacted by agriculture and urbanization (site). Four invertebrate PIs will be further

considered for indicator development. The

richness of snail taxa (GASR) is positively associated with forest and wetland cover from 1,000 to

5,000 m.
49

Montana: Developing Wetland Bioassessment

Protocols To Support Aquatic Life Beneficial

Use-Support Determinations

Contact Randall S. Apfelbeck

Organization Montana Department of Environmental Quality

2209 Phoenix Avenue

P.O. Box 200901

Helena, MT 59620-0901

Phone (406) 444-2709

E-mail rapfelbeck@state.mt.us

Website http://www.deq.state.mt.us/ppa/mdm/Wetlands/Lakes&Wetlands_Index.asp

Purpose

Montana’s wetland research objectives are to:

 Determine the status and trends in wetland water quality.

 Acquire an understanding of how climate, hydrologic controls, and geomorphic settings influence wetland biological communities for the

development of successful biocriteria.

 Develop biological measurements that could be

used in developing biocriteria to define the extent and degree of anthropogenic impacts to

wetland water quality.

 Develop an integrated assessment of wetland,

streams and lakes within a watershed.

 Report on aquatic health at the watershed level

through the development of landscape assessment tools.

 Develop rapid field assessment protocols to

evaluate aquatic ecological conditions by using

indicators and best professional judgement.

Wetland Type

 Variety of wetland types

Assemblages

 Algae

 Macroinvertebrates

 Vegetation

 Amphibians and aquatic reptiles

Status

 Ongoing, revising analytical methods

Project Descriptions

1992

Montana Department of Environmental Quality

(DEQ) began developing wetland biological criteria. At that time, there was little information concerning the status or trends of the water quality of

Montana’s wetlands. Furthermore, Montana’s

water quality standards were developed to protect

the beneficial uses (e.g., aquatic life) of lakes; river
50

and stream wetlands were not considered State

waters when Montana’s water quality standards

were developed. Since 1992, Montana has had an

ongoing program to develop bioassessment protocols and water quality standards that will more adequately evaluate and protect the aquatic life that

live in wetlands.

In 1998, as a result of a TMDL lawsuit, Montana

DEQ shifted focus from development of biological

criteria to the development of guidelines for making

beneficial use decisions that apply to all Montana

waterbodies. Montana’s guidelines for making beneficial use decisions can be found at http://

nris.state.mt.us/wis/tmdlapp/pdf2002/

Appendix_A.pdf .

1998

In conjunction with Montana DEQ’s research program, Montana State University (MSU) designed

a study that focused on development of vegetation

biocriteria for western Montana depressional wetlands (Borth 1998). The focus on vegetation

biocriteria is key in Montana because wetland vegetation is easier to assess than macroinvertebrates

or diatoms for depressional wetlands that are seasonally dry. The MSU study sampled vegetation

and also macroinvertebrates and diatoms for 24 depressional wetlands with similar climate, hydrology,

and water chemistry. The research included sampling across three levels of human disturbances—

minimally impacted, slightly impacted, and moderately impacted. The study also involved two anthropogenic impairments—dryland agriculture and

grazing.

2000

Researchers from the University of Montana conducted a study to determine the effects of natural

variability on the use of macroinvertebrates as

bioindicators of disturbance in intermontane depressional wetlands in northwestern Montana (Ludden

2000). Their study design included collection of

macroinvertebrate samples and physiochemical data

from 15 pristine and 6 disturbed intermontane prairie potholes. Their study also included analysis of

macroinvertebrate samples that were previously

collected by Borth (1998). The researchers collected macroinvertebrate samples from across three

seasons and from three wetland zones. They used

multivariate detrended correspondence analysis to

ordinate the raw macroinvertebrate data and

physiochemical variables as secondary matrices to

establish vectored biplots of correlation. Candidate metrics were analyzed using univariate analysis. They determined that no environmental variables were strongly correlated with the

macroinvertebrate data, and many of the metrics

varied across wetland zones and across seasons.

Nevertheless, 35 candidate metrics were able to

discriminate between minimally and highly disturbed

sites. These wetlands were also sampled intensively

to develop and test a model for assessing depressional wetlands using the hydrogeomorphic (HGM)

functional assessment approach.

2002

Montana DEQ initiated the development of a

comprehensive watershed monitoring and assessment program. The comprehensive program will

be developed to determine the causes, effects, and

extent of pollution to aquatic resources (including

wetlands) and for developing pollution prevention,

reduction, and elimination strategies.

The comprehensive program has three components: landscape, site-specific, and rapid assessment. First, a landscape-level process to evaluate

and rank wetlands and watersheds for protection

and restoration is critical to maximize the use of limited financial and management resources. This process will use existing digital data and evaluate a range

of landscape impacts using a Geographic Information System. Second, Montana will continue to

develop site-specific assessment protocols such as

biocriteria (i.e., for assessing amphibians, vegetation, algae, and macroinvertebrate assemblages).

This information is important for managers of

Montana’s water resources for making informed

watershed management decisions. Development
51

of site-specific wetland biological, physical, and

chemical assessment tools will be used to evaluate

aquatic life beneficial use support to determine impairment as per section 303(d) of the Clean Water

Act. Third, like most States, Montana has very

limited resources for assessing wetland water quality and aquatic conditions for 305(b) purposes. For

this reason, Montana will develop an assessment

approach that is truly rapid and cost effective. A

rapid assessment component will be developed

based upon field data collected, tested and refined

during this study, best professional judgment and

information gathered from a literature search.

The comprehensive watershed monitoring and

assessment program will include a probabilistic study

design to assess the ecological condition of depressional and riverine wetlands and streams. Three

pilot subbasins (fourth code HUCs) will be selected.

The pilot subbasins will represent Rocky Mountain, Intermountain Valleys and Prairie Foothills, and

Plains ecoregions (Figure 11).

Within each pilot subbasin, three sixth-code HUCs

(fifth-code HUCs for the Middle Milk subbasin)

will be randomly selected from each of three disturbance strata (high, medium, and low) determined

from landscape-scale assessments. For vegetation,

soils, water chemistry, macroinvertebrates, diatoms,

and land use, a total of six wetlands (three riverine

and three depressional) will be monitored in each

watershed. A total of 27 watersheds (9 in each

subbasin) and 162 wetlands (3 riverine and three

depressional in each watershed) will be studied. For

the determination of the detection/nondetection of

amphibians and aquatic reptiles, all standing water

bodies identified in each watershed will be surveyed.

Note: the remainder of this case study will describe in more detail the study initiated in 1992.

Study Design

The original Montana DEQ study was designed

in 1992 and involved sampling 80 wetlands

throughout Montana during 1993 and 1994. The

bioassessment project included collection of

macroinvertebrate and diatom samples from wetlands in all ecoregions of Montana (Apfelbeck

2001).

Montana DEQ’s approach to developing

biocriteria involved several study designs aimed at

developing tools to help detect human influence on

wetland water quality. The original study was designed in 1992 and involved sampling 80 wetlands

Figure 11: Montana ecoregions and pilot watersheds.
52

throughout Montana during 1993 and 1994 (Figure 12). The study design included collection of

samples that represent the wetland’s

macroinvertebrate (e.g., aquatic insects) and diatom (algae) communities. Sampling methods were

designed so that 1-2 hours was sufficient for the

data to be collected in the field for each wetland.

Samples of each wetland’s water column, sediment,

and macroinvertebrate and diatom communities

were collected. Water-column and sediment

samples were collected to document impairments

and for classification purposes.

Wetland Classes: (referred to by number in the above figure)

Class 1 Dilute Closed Basins and Headwater Wetlands of the Rocky Mountain Ecoregion

Class 2 Riparian Wetlands of the Rocky Mountain amd Intermountain Valley Ecoregions

Class 3 Groundwater Recharge Closed Basin Wetlands

Class 4 Riparian Wetlands of the Plains Ecoregion

Class 5 Alkaline Closed Basin Wetlands

Class 6 Saline Wetlands

Class 7 Surface Water Supported Closed Basin Wetlands

Class 8 Ephemeral Wetlands

Class 9 Open Lake Wetlands of the Plains Ecoregion

Class 10 Open Lake Wetlands of the Rocky Mountain and Intermountain Valley Ecoregions

Figure 12: Ecoregions and sampling locations by wetland class.

Ecoregions of Montana
53

The sites were classified using ecoregions and

hydrogeomorphology, and several of the wetland

classes were further delineated using water-column

chemistry variables. A representative number of

wetlands from the following Omernik ecoregions

were sampled: Rocky Mountains, Intermountain

Valleys and Prairie Foothills, Glaciated Plains, and

Unglaciated Plains Ecoregions. To reduce seasonality, all wetlands within the same ecoregion were

sampled during similar time periods. Wetlands in

the Glaciated Plains Ecoregion were sampled from

early April through mid-June, wetlands of the Intermountain Valleys and Prairie Foothills Ecoregion from

mid-June until early August, and wetlands of the

Rocky Mountain Ecoregion from early July through

September.

The classification framework was developed by

sampling the full spectrum of wetland types in Montana. The study was designed such that approximately 75% of the sites were reference and 25%

were impaired. This approach was useful because

it allowed Montana DEQ to determine the reference condition of a wide variety of wetland types.

Also, the design provided the opportunity to test

the ability of the biological measurements to detect

water quality impairment.

Anthropogenic impacts such as irrigation or logging were included in the study design. If anthropogenic activities such as dryland agriculture, irrigation, feedlots, grazing, silviculture, road construction, hydrologic manipulation, urban runoff, wastewater, mining, and oil and natural gas production

occurred in the wetland’s watershed, the wetland

was considered impaired. Wetlands for sampling

were selected on the basis of many variables, including availability of historical data, special interests by other entities, cooperation by landowners,

and accessibility.

In order to classify or document impairment, a

hydrogeologist for the Montana Natural Heritage

Program (MNHP) assisted Montana DEQ in developing a wetland classification system through

summarizing and interpreting the physical and chemical data. Using topographic maps, field observations, and information gathered from land management agencies, geomorphic characteristics were interpreted and a hydrogeomorphic database developed. Maps for each wetland were completed using a Geographic Information System (GIS). Map

features included hydrologic delineations, land management areas, counties, cities, major transportation corridors, wetland watershed boundaries, and

sampling locations. Visit Montana DEQ’s website

to get more detail on the types of wetland classes

and for photographs of each type: http://

www.deq.state.mt.us/ppa/mdm/Wetlands/

classification.asp.

Sampling Methods: Diatoms

Montana DEQ collected diatoms as composite

grab samples. The algae were identified to the lowest

taxonomic level possible. Samples were collected

using a 250-mL plastic container and then preserved

with Lugol’s solution. Samples were collected from

a location determined to best represent the wetland. These locations were restricted to areas that

were easily accessible when wearing hip boots.

Sampling was done from April through September.

Each site was sampled once.

Analytical Methods: Diatoms

The multivariate approach was used to analyze

wetland diatom communities. Multivariate analysis

is a statistical approach used by biologists to determine relationships among biota such as diatoms or

macroinvertebrates, and environmental variables

such as water-column chemistry. The multivariate

approach to investigate relationships between Montana wetland diatom assemblages and environmental variables (mostly water-column chemistry) was

detrended canonical correspondence analysis

(DCCA) and two-way indicator analysis
54

(TWINSPAN). Clusters of diatoms with similar

composition were graphically displayed using

DCCA. Vectors were displayed and labeled to

illustrate the relationship between diatom assemblages and environmental variables. Longer vectors show us a stronger correlation among diatom

assemblages and environmental variables (Figure

13). Envelopes were used to graphically enclose

all reference sites using the wetland class delineations (Figure 14). The Academy of Natural Sciences of Philadelphia (ANSP) performed the

subsampling, digestion, and mounting of the diatoms (see Charles et al. 1996).

Figure 13: Diatom data related

 to abiotic factors using

Detrended Canonical

Correspondence Analysis.

Figure 14: Clusters of wetlands

based on diatom data.
55

Sampling Methods:

Macroinvertebrates

Montana DEQ collected macroinvertebrates using a 1-mm mesh D-net in a sweeping motion.

Macroinvertebrates were collected from all microenvironments in a sampling location. These locations

were restricted to areas that were easily accessible

when wearing hip boots and that best represented

the wetland. Samples were composited with associated materials such as vegetation and sediment

and then preserved with ethanol in a 1-L plastic

container. An effort was made to collect 300 organisms from each location using a consistent

method of collection. To ensure preservation,

sample bottles were refreshed with ethanol several

days after collection. Sampling was done once per

site from April through September.

Analytical Methods:

Macroinvertebrates

The multimetric approach was used to evaluate

wetland macroinvertebrate communities. The

multimetric approach incorporates many attributes

into the assessment process and has the ability to

integrate information from the biological communities to provide an overall indication of biological

condition or ecological health.

The project contractor assisted Montana DEQ in

developing wetland macroinvertebrate multimetric

indices. The proposed metrics and associated environmental data were evaluated in an attempt to

develop an understanding of ecological relationships,

to test each proposed metric’s ability to predict

various anthropogenic stressors, and to test redundancy. Table 3 lists some of the proposed metrics

used for macroinvertebrates and Figure 15 shows

macroinvertebrate index values for several classes

of wetlands.

The macroinvertebrate samples are subsampled

and sorted by contractors using a gridded sorting

pan. A subsampling of 200 organisms is performed

for analysis. All individuals are counted when there

are less than 200 organisms in the sample. Organisms are identified to the lowest taxonomic level (at

least genus if possible). The taxonomic level of identification is standardized using the Montana Stream

when possible. Amphipoda are identified by species. Common, easily identified midge taxa are identified using a dissecting microscope equipped with

25H oculars. Midge taxa that require slide mounting are cleaned with warm water solution of 10%

KOX, rinsed in distilled water followed by 95%

ethanol, and mounted on slides in Euperal. If fewer

than 30 individuals are to be mounted, all individuals are mounted. If there are more than 30 midge

larvae, at least 10% of each morphotype are

mounted. Mounted midges are identified using a

Zeiss Axiolab phase contrast compound microscope

or equivalent.

The contractor identified the organisms in the

wetland samples and standardized the taxonomic

level of identification based on Montana Stream

Protocols. Several taxa were eliminated from consideration for metric development, as they were

determined to be nonbenthic taxa or semiaquatic

surface dwellers and considered uninformative for

reflecting water quality. These taxa included

Gerridae, Collembola, Dytiscidae, Hydrophilidae,

Ostracoda, Anostraca, Copepoda, Cladocera,

Notonectidae, and Corixidae.

Other Parameters: Water

Chemistry and Sediment

Water-column and sediment samples were collected to document impairments and for classification purposes. Each sample was collected from a

location determined to best represent the wetland.

These locations were restricted to areas that were

easily accessible when wearing hip boots. Field

chemical measurements, observations, and photographs were recorded at each location.
56

Table 3: Proposed metrics, proposed metric calculations, and score

calculations used for developing wetland macroinvertebrate indices

Lessons Learned

 We found that diatoms and macroinvertebrates

were most useful for evaluating the biological

integrity of perennial wetlands with open-water

environments that had relatively stable water

levels and were not excessively alkaline or saline.

 We concluded that multivariate analysis was a

useful tool for developing a wetland classification system and that hydrogeomorphology and

ecoregions were practical approaches to classifying wetlands for the development of

biocriteria.

 We determined that both the multimetric and

multivariate techniques were valuable for developing wetland biocriteria.

 In most cases, the multimetric and multivariate

approaches that we used to assess the

macroinvertebrate and diatom communities both

identified the same wetlands as impaired.

 Two wetland types in the arid west (including

Montana) are difficult to classify. Wetlands such

as potholes are highly complex and difficult to

classify because of both spatial and temporal

variability. For these wetlands, the hydrology,

water chemistry, and biology can change dra-
57

Figure 15: Macroinvertebrate index scores for wetlands

in several classes.
58

matically throughout a season or from year to

year as a result of climatic change. For example,

the biological community of a wetland often

changes through an increase in salinity or a decrease in water content caused by drought.

 Aquatic macroinvertebrate and diatom

biocriteria did not appear to be very useful for

detecting impairment in wetlands that usually

lack surface water. Vegetation biocriteria are

likely to be the most appropriate for assessing

the biological conditions of these wetland types.

 A more quantitative approach is needed for the

measurement of wetland physical disturbances

if we are to link physical disturbances to

changes in biological communities.

Note: Montana DEQ has developed a set of proposed metrics, proposed metrics calculations, and

score calculations used for developing wetland

macroinvertebrate indices. These are included in

Table 3.
59

North Dakota: Wetland Bioassessment

Protocols for Making Aquatic Life Beneficial

Use-Support Determinations

Contact Mike Ell

Organization North Dakota Department of Health

Division of Water Quality

1200 Missouri Avenue

P.O. Box 5520

Bismarck, ND 04333

Phone (701) 328–5214

E-mail mell@state.nd.us

Purpose

The primary purpose of North Dakota’s wetland

bioassessment program is to develop wetland water quality standards for North Dakota. This currently involves developing biological community

metrics and an Index of Biological Integrity (IBI)

for temporary and seasonal depressional wetlands.

A secondary project goal is to compare the IBI

with hydrogeomorphic (HGM) functional assessments.

Wetland Type

 Depressional wetlands

Assemblages

 Algae

 Macroinvertebrates

 Vascular plants

Status

 Analyzing data, developing IBIs, and expanding project

Project Description

North Dakota Department of Health (NDDH)

initiated its wetland bioassessment program in 1993

as a component of the State’s strategy to develop

wetland water quality standards. NDDH began field

sampling in 1995 and is currently analyzing data

collected from 1995 through 1999 to develop assemblage metrics, IBIs for the assemblages, and

ultimately biocriteria. The NDDH wetland sampling plan includes vascular plants,

macroinvertebrates, and algal assemblages.

Development of a North Dakota sampling

protocol for the State’s wetland types is ongoing

through cooperative work agreements with North

Dakota State University, Departments of Animal

and Range Sciences and Zoology.

Study Design

North Dakota’s study design has continued to

expand in scope since the project was initiated. Initial field sampling began in 1995 and 1996 with 13

“least disturbed” temporary and seasonal wetlands.

In 1997, the number and range of wetlands was

expanded to 20 temporary and seasonal wetlands.

Four of the original 13 “reference” wetlands sampled
60

in 1995 and 1996 were sampled in 1997 along with

16 new wetlands. These wetlands captured a larger

range of disturbance categories.

In 1998, the number of wetlands again expanded.

An additional 16 temporary and seasonal wetlands

were added, bringing the total number of wetlands

sampled to 36 in 1998 and the total number of reference wetlands to 6. The 16 wetlands added in

1998 were part of an HGM classification project.

These wetlands were added to facilitate a comparison of the two assessment methods. In addition to

the 16 HGM wetlands, NRCS personnel conducted

an HGM assessment for the 20 wetlands selected

by the Department.

During the 1999 sampling season, a second set of

30 temporary and seasonal depressional wetlands,

not sampled previously, were sampled for vascular

plants. Macroinvertebrates and algae were not

sampled in 1999.

Since the inception of the project, temporary and

seasonal wetlands have been the focus of IBI development efforts. Further, all wetland study sites

have been within the Northern Glaciated Plains and

Northwestern Glaciated Plains ecoregions (Figure

16), a region commonly referred to as the “Prairie

Pothole Region.”

Sampling Methods:

Macroinvertebrates

NDDH currently uses only the “sweep” or jab

net method for macroinvertebrate sampling; however, in the past, activity traps have also been used.

Sampling consists of two site visits in June. During

each site visit two samples are collected with the

emergent vegetative zone.

Sweep net

The method employs a “D” frame net (0.6-mm

mesh size). At each site within the wetland, the

substrate (i.e., wetland vegetation and benthos) is

“jabbed” for a distance of 1 m. The disturbed area

is then swept two more times to ensure a representative sample of macroinvertebrates is collected.

Each sample collected within the wetland will be

placed in a shallow pan where any excess debris or

water can be removed. The “cleaned” sample is

placed in a jar a preserved with buffered formalin

to a concentration of 10% by volume.

Analytical Methods:

Macroinvertebrates

In the laboratory each sample is washed through

a 0.6-mm sieve to remove the preservative and

cleaned to remove excess debris. The cleaned

sample is placed in a shallow white pan divided in

quadrants of equal size. Because each sample typically contains a large number of organisms, it is necessary to subsample. A subsample is obtained by

randomly selecting a sample quadrant and counting

and identifying the organisms in that quadrant. Additional quadrants are selected at random and organisms counted and identified until a subsample of

300 organisms is counted. For samples with less

than 300 organisms, every quadrant is sampled. All

organisms are identified to the lowest taxonomic

level practical.

Sampling Methods: Algae

(Phytoplankton)

Phytoplankton samples are collected just below

the water’s surface in the middle or deepest area of

the wetland basin. Each wetland is sampled two

times in June at the same time macroinvertebrate

samples are collected. The sample is collected by

submerging a clean 200-mL sample bottle just below the water’s surface and filling it. The sample is

preserved in the field with M3 fixative to a concentration of 2% by volume.
61

Figure 16: Level IV ecoregions of North Dakota.
62

Analytical Methods: Algae

(Phytoplankton)

Identification and enumeration of phytoplankton

is made using the phase-contrast inverted microscope method (Utermohl 1958). Sample analysis

is conducted from a sample aliquot ranging from 2

to 15 mL, depending on turbidity. Phytoplankton

greater than 20 micrometers in diameter are enumerated at H125 magnification. (Note: At H125

the entire bottom of the chamber is in view.) Following enumeration at H125 smaller algae are

counted at H1,250. At least 250 cells of the most

numerous algae are counted using the strip count

method at H1,250 magnification (APHA 1989).

Cell volumes are estimated for dominant taxa by

measuring cell dimensions of 50 to 100 individuals

using the closest geometric formulas of Wetzel and

Likens (1991) and Tikkanen (1986). For rare taxa,

volume estimates are made from fewer than 50 cell

measurements. Diatoms will be identified after

clearing in 30% hydrogen peroxide and mounting

in Hyrax Mounting Medium.

Sampling Methods:

Vascular Plants

Working in cooperation with NDSU, the Department is evaluating three methods to sample the vascular plant community. The first method is more

qualitative in nature. It involves a simple inventory

of plant species present within each wetland zone,

low prairie, wet meadow, and shallow marsh. The

other two methods, termed the point and quadrat

methods, are quantitative. The inventory is conducted each time the wetland is visited. The point

and quadrat methods are done once each, usually

in July or August.

In the point method, 200 points are evenly stratified in each wetland zone and the nearest plant species recorded. The quadrat method involves placing 15 1-m2

 quadrats evenly throughout each wetland zone. Each species is recorded within the

quadrat and a Daubenmire cover class is recorded.

With both methods, a secondary species list is made

for species encountered, but not sampled.

Note: For specific details on abiotic sampling

methods for nutrients, trace elements, general water parameters, and sediment chemistry, directly refer

to North Dakota Department of Health.

Lessons Learned

NDDH has made a great deal of progress within

the past 5 years. Our experience, however, has

not been without problems and mistakes. As mentioned previously, the first 2 years of the program

focused solely on “least impaired” or reference condition wetlands. Although beneficial in testing sampling methods, the lack of a disturbance gradient in

the study design did allow for the testing of attributes

and the selection of metrics. Therefore, beginning

in 1997 the NDDH chose wetlands across a disturbance gradient, including both reference wetlands

and degraded wetlands.

NDDH has also found it beneficial to stratify wetlands based on ecoregion and wetland class. This

minimizes the amount of variation in the biological

assemblage and allows more sensitivity in the response of the metrics to the disturbance gradient.

Current IBI development efforts are focusing on

temporary and seasonal depressional wetlands

within the Northern Glaciated Plains and Northwestern Glaciated Plains ecoregions. In 2000 the

NDDH will be cooperating in two projects to develop wetland IBIs for semipermanent depressional

wetlands and for floodplain wetlands along the Missouri River.
63

Ohio: Developing IBI Assessment Methods

for Water Quality Standards and

Regulatory Decisions

Purpose

 Test and develop biological criteria for wetlands

using vascular plants, macroinvertebrates, and

amphibians as indices of biological integrity

(IBIs) for eventual adoption into the State’s

water quality standards

 Use results from IBIs to calibrate the Ohio Rapid

Assessment Method for Wetlands to support

regulatory decisionmaking under the State’s

wetland antidegradation rule, which requires that

wetlands be assigned to one of three categories based on the wetland’s quality and functionality.

Wetland Type

 Depressional wetlands

Assemblages

 Algae

 Macroinvertebrates

 Vascular plants

Contact John J. Mack

Organization Ohio EPA

Division of Surface Water

122 South Front Street

P.O. Box 1049

Columbus, OH 43216-1049

Phone (614) 644-3076

E-mail john.mack@epa.state.oh.us

Status

 Refining metrics and developing IBIs

Project Description

The initial objective of this study is to provide the

reference data needed to implement the wetland

water quality standards and wetland antidegradation

rule. The pilot metrics developed from this study

should enable Ohio wetlands to be assigned to one

of the three regulatory categories. Generally, the

study objectives are as follows:

1. Develop pilot biological metrics that may be

used to evaluate the function and ecological integrity of a wetland. These metrics will be based

on the vegetation, macroinvertebrate, and amphibian data, and will form the basis for wetland biocriteria.

2. Identify and describe reference wetlands in

Ohio’s four main ecoregions; Eastern Cornbelt

Plains, Erie/Ontario Drift and Lake Plain, Huron-Erie Lake Plain, and Western Allegheny

Plateau. These reference wetlands will be used

to develop biocriteria and also as “goals” for

wetland mitigation projects.
64

3. Continue to assess whether the Ohio Rapid

Assessment Method correlates well with the

more in-depth measures of wetland quality, and

to test and refine breakpoints between the wetland categories.

4. Begin to assess the sensitivity of different methods in evaluating the relationship between wetland quality and the degree of disturbance

The State of Ohio has well-developed biological

criteria (or biocriteria) for streams, such as the Invertebrate Community Index (macroinvertebrates),

the Index of Biological Integrity (fish), and the Modified Index of Well Being (fish) (Ohio EPA 1988a,b

and 1989). These indices are codified in Ohio

Administrative Code Chapter 3745-1. Until recently, however, surface waters of the State that

are jurisdictional wetlands were only generically

protected under Ohio’s water quality standards.

On May 1, 1998, the State of Ohio adopted wetland water quality standards and a wetland

antidegradation rule. These wetland quality standards developed narrative criteria for wetlands and

created the “wetland designated use.” All jurisdictional wetlands are assigned the “wetland designated

Figure 17: Ecological regions of Ohio and Indiana

(from U.S. EPA 1995).

Ecoregions** of Ohio and Indiana and Neighboring States
65

use.” The State of Ohio did not attempt at this

time to identify multiple wetland functions as wetland uses, because of the lack of data to develop

quantitative water quality criteria for wetlands.

However, the development of such biocriteria is the

ultimate goal and the primary thrust of this project.

The key part of Ohio’s current regulatory program for wetlands is found in the wetland

antidegradation rule. The wetland antidegradation

rule categorizes wetlands based on their functions,

sensitivity to disturbance, rarity, and irreplaceability, and scales the strictness of avoidance, minimization, and mitigation to a wetland’s category.

Three categories were established: Category 1

wetlands with minimal wetland function and/or integrity; Category 2 wetlands with moderate wetland function and/or integrity; and Category 3 wetlands with superior wetland function and/or integrity. In order to implement the wetland standards

and antidegradation policy, wetlands must be assessed on their relative quality. Ohio EPA has developed a draft Ohio Rapid Assessment Method.

The Ohio Rapid Assessment Method has proved

to be a fast, easy-to-use procedure for distinguishing between wetlands of differing quality. However, it does not and was not intended to substitute

for direct, quantitative measures of wetland function (i.e., biocriteria).

Ohio began development of sampling methodologies and began sampling reference wetlands for

biocriteria development in 1996. To date, Ohio

has sampled 56 wetlands located primarily in the

Eastern Cornbelt Plains Ecoregion located in central and western Ohio. These wetlands have included depressional emergent, forested, and scrubshrub wetlands; floodplain wetlands; fens; kettle

lakes; and seep wetlands. The wetlands being studied span the range of condition from “impacted”

(i.e., those that have sustained a relatively high level

of disturbance) to “least-impaired” (i.e., the best

quality sites available).

On the basis of results to date (see Fennessy et

al. 1998a,b; Mack et al. unpublished data), Ohio’s

research supports the use of vascular plants,

macroinvertebrates and/or amphibians as biological metrics in wetlands, and also the continued use

and development of the Ohio Rapid Assessment

Method as a rapid assessment tool.

This work has been funded since 1996 by several

EPA Region 5 Wetland Program Development

Grants.

Study Design

Fifty-seven wetlands were sampled during the

1996, 1997, 1998, and 1999 field seasons. The

first 2 years of data laid the groundwork for standardizing sampling methodologies, classifying wetlands, identifying potential attributes, and developing metrics using vascular plants, amphibians, and

macroinvertebrates.

In 1996, Ohio EPA monitored a series of riparian

forested across a gradient of disturbance (i.e., least

impacted to impaired) (Fennessy et al. 1998b).

Estimates of the relative level of disturbance were

made on a scale of 1 (most disturbed) to 10 (least

disturbed) based on visual evidence of disturbances,

review of aerial photographs of the wetland and the

surrounding area, and interviews with staff from the

Natural Resource Conservation Service and/or the

landowner. In 1996 and 1997, Ohio EPA monitored 21 forested and emergent depressional wetlands. Relative disturbance was evaluated using a

tiered flow chart to assign a relative disturbance

score and also with the score from the Ohio Rapid

Assessment Method (Fennessy et al. 1998a, Figure 2.2).

Ohio EPA found a good correlation between the

scores of the Ohio Rapid Assessment Method score

and level of disturbance a wetland site has experienced. Higher ORAM scores correlate well with
66

lower levels of disturbance based on our model, as

do lower ORAM scores with disturbed sites. In

1999, the ORAM score of the site was used as a

measure of the level of disturbance. So far, this

appears to be a highly effective “x-axis” disturbance

gradient for the development of IBIs for wetland

plants.

Reference wetlands are sites or data sets from

sites that typify a class of wetlands within a relatively homogeneous physiographic region. Reference sites should include wetlands that have been

degraded or disturbed. Site selection in this study

is made using an ecoregional approach and to reflect a gradient of disturbance (i.e., least-impacted

to impaired).

Reference sites are selected such that relatively

similar proportions of low-, medium-, and high-disturbance sites are sampled. To date, almost all the

wetlands studied by Ohio EPA have been located

in the Eastern Cornbelt Plains (ECBP) ecoregion.

For the years 2000 and 2001 field seasons, Ohio

EPA will be studying reference sites in the Erie

Ontario Lake Plain (EOLP) ecoregion of northeastern Ohio.

Sampling Methods and Analysis:

Macroinvertebrates and

Amphibians

Below is a detailed description of the sampling

methodology and analysis process used by Ohio

EPA for macroinvertebrates and amphibians. Also

included are the lessons learned for sampling these

assemblages.

Amphibian and macroinvertebrate taxa were selected as potential indicators of wetland condition.

On the recommendation of many field professionals, experienced in amphibian monitoring, funnel

traps were used. The funnel traps also proved to

be extremely effective at sampling wetland

macroinvertebrate communities. Therefore, the

same sampling protocols are used for both amphibians and macroinvertebrates in Ohio EPA’s study.

Funnel trap

The funnel traps used in this project have cylinders constructed of aluminum window screen and

funnel ends made from fiberglass window screen.

The funnel traps are similar in design to commercially available minnow traps. The cylinders are 18

inches long and 8 inches in diameter. The two funnel ends are attached to the cylinders and begin 8

inches in diameter and taper inward 5 inches to a

13⁄4-inch opening. A string handle that runs from

end to end is attached to the two seams where the

cylinder and funnels ends join.

Plate 7: Impaired wetland.

Photo: Mick Micacchion

Plate 8: Least-disturbed wetland.

Photo: Mick Micacchion
67

To date, Ohio sampling data have come from depressional wetland systems in the Eastern Corn Belt

Plains (ECBP) ecoregion. The Eastern Corn Belt

Plains ecoregion spans most of the western half of

Ohio and accounts for about 40% of Ohio’s land

mass. Within the ECBP, depressional study wetlands that demonstrated a gradient of disturbance

levels, from least impacted to greatly disturbed, were

selected.

Selected wetlands are sampled for amphibians and

macroinvertebrates three times during the year

(early, middle, and late spring). An early spring

sample is collected in the period from late February

to late March. This early spring sample run allows

sampling of adult Ambystomatid salamanders and

early spring macroinvertebrates (e.g., fairy shrimp)

that are only present in wetlands for a limited time.

Adult salamanders enter wetlands to breed in early

spring following the first few warm, rainy nights of

late winter and early spring. The actual timing of

their arrival in Ohio is highly weather dependent and

varies greatly by year and location. A second

sample is collected during the month of April in order to collect adult frogs entering wetlands to breed,

as well as amphibian larvae already present, and to

sample for macroinvertebrates. Mosquito larvae,

an important prey item for many predators, are

abundant in April in Ohio wetlands. The final sampling is conducted between May 15 and June 15.

Salamander larvae and frog tadpoles are collected,

in addition to the resident macroinvertebrates. This

last sampling run occurs late enough in the breeding

cycle to allow collection of larvae from all breeding

amphibians. However, it is still early enough that

even in drought years temporary wetlands will not

have dried up.

Generally, 10 funnel traps are installed at each

wetland. Prior to installing the first funnel trap, the

perimeter of the area where standing water is

present is measured using a hip chain or by pacing.

The total perimeter length is then divided by 10,

and 10 funnel traps are installed uniformly around

the perimeter of the wetland at intervals of 10% of

the total perimeter distance. Each funnel trap location is permanently marked with flagging for use

throughout the sampling season. The funnel traps

are installed on the bottom at a location deep enough

to submerge the trap. The traps are left in the wetland 24 hours to ensure unbiased sampling for animals with diurnal and nocturnal activity patterns. The

design of the traps allows them to collect any amphibians and macroinvertebrates that swim or crawl

into the funnel openings.

Upon retrieval, the traps are emptied by inverting

one of the funnels and dumping and shaking the

contents into a white sorting pan. Organisms that

can be readily identified in the field are counted and

recorded in the field notebook and released. The

remaining organisms are transferred to a 1-L plastic bottle and preserved with 70% ethanol. The

contents of each trap are kept in separate, clearly

marked bottles for individual analysis in the laboratory. If large numbers of amphibians are kept for

identification in the lab, the samples are transferred

to formalin for long-term storage. All organisms

collected are identified in the lab using appropriate

keys and the results are recorded.

Dipnet

Qualitative collections are made concurrently with

funnel trapping at each wetland once during each of

the three sampling periods. Qualitative sampling

involves the collection of amphibians and

macroinvertebrates from all available natural wetland habitat features. This is achieved by using triangular ring frame 30-mesh dipnets and manual

picking of substrates with field forceps. The goal is

to compile a comprehensive species/taxa list of

macroinvertebrates and amphibians at the site. A

minimum of 30 minutes is spent collecting the qualitative sample. Sampling continues until the field crew

determines that further sampling effort will not produce new taxa. At least one specimen of all taxa

collected during the qualitative sampling is preserved

in a jar of ethanol for positive identification in the

laboratory.
68

Hester-Dendy artificial substrate sampler

Five Hester-Dendy (HD) artificial substrate samplers were tied to the top of a concrete block and

placed in wetlands where they remained submerged

for 6 weeks. The samplers were collected and preserved in formalin. All the macroinvertebrates colonizing the samplers were counted and identified.

Analytical Methods:

Macroinvertebrates and

Amphibians

Macroinvertebrates were identified to genus or

species. Amphibians were identified to species except for some small salamander larvae identified to

genus. Each funnel trap collection was analyzed

individually so that location-specific information was

not lost by pooling all samples from a site.

The number of individuals collected in the traps

was divided by the number of hours trapped to give

a relative abundance consisting of number of individuals per trap hour. Results from the different

wetland study sites were examined for faunal differences in distribution and abundance. Analysis of

the data for potential biological indicators of human

disturbance is under way.

Lessons Learned for

Macroinvertebrates and

Amphibians

 Funnel traps consistently collected an average

of 10 more macroinvertebrate taxa than qualitative sampling using dipnets. Funnel traps were

much more effective in sampling amphibians and

fish than sampling with dipnets.

 Qualitative sampling collected somewhat more

Mollusca and Chironomidae taxa than did funnel traps.

 Funnel traps collected more leech taxa, Hemiptera taxa, Coleoptera taxa, Odonata taxa, and

Crustacea taxa than did qualitative sampling.

 Hester-Dendy artificial substrate samplers were

ineffective for sampling most wetland

macroinvertebrates, except oligochaetes,

Chironomidae, and Mollusca.

 A 24-hour sampling period for funnel traps is

preferred as it allows for the collection of nocturnal species that are infrequently collected by

daytime sampling methods.

A single wetland often has several vegetation

classes. Even if only three main classes are identified (forested, scrub-shrub, and emergent), the

wetlands included for study can exhibit multiple combinations. For example, of the 20 wetlands studied

in Fennessy et al. (1998a), 6 combinations of vegetation classes were found: emergent, emergent/

scrub-shrub, forested, forested/emergent, forested/

emergent/scrub-shrub, and forested/scrub-shrub.

Thus, a sampling method should be flexible enough

to account for horizontal and vertical variation in

vegetation.

After testing a transect-quadrat method, Ohio EPA

has adapted the method used by the North Carolina Vegetation Survey as its standard vegetation

sampling method (Peet et al. 1998). This is a flexible, multipurpose sampling method that can be used

to sample such diverse communities as grass- and

forb-dominated savannahs, dense shrub thickets,

forest, and sparsely vegetated rock outcrops, and

has been used at more than 3,000 sites over 10

years as part of the North Carolina Vegetation Survey. This method is appropriate for most types of

vegetation, flexible in intensity and time commitment,

compatible with other data types from other methods, and provides information on species composition across spatial scales. It also addresses the

problem that processes affecting vegetation composition differ as spatial scales increase or decrease

and that vegetation typically exhibits strong

autocorrelation (Peet et al. 1998). Peet et al. (1998)

state, “Our solution to the problems of scale and

spatial auto-correlation is to adopt a modular approach to plot layout, wherein all measurements are
69

made in plots comprised of one or more 10x10-m

quadrats or “modules” (100 m2 = 1 are = 0.01 hectare). The module size and shape were chosen to

provide a convenient building block for larger pots,

and because a body of data already exists for plots

of some multiple of this size. The square shape is

efficient to lay out, ensures the observation is typical for species interactions at that scale of observation, and avoids biases built into methods with distributed quadrats or high perimeter-to-area ratios”

(Peet et al. 1998, p. 264). Basically, the method

employs a set of 10 modules in a 20H50-m layout.

Within the site to be surveyed, these 20H50-m grids

are located such that the long axis of the plot is

oriented to minimize the environmental heterogeneity within the plot.

Once the plot is laid out, all species within the plot

are identified, an aggregate wood stem count is made,

and cover is estimated at the 0.1-hectare scale. In

addition, four 10H10-m modules are intensively

sampled in a series of nested quadrats. Within these

“intensive” modules, species cover class values and

woody stem tallies are recorded for each module

separately and for each nested quadrat separately.

In effect then, the method proposed by Peet et al.

incorporates use of reléves found in the BraunBlanquet methodology inasmuch as the length,

width, orientation, and location of the modules are

qualitatively selected by the investigator on the basis of site characteristics; however, within the modules, standard quantitative floristic and forestry information is recorded, such as density, basal area,

cover, etc.

Once the location of the plot or plots has been

selected, the primary purpose of the vegetation survey is to obtain a comprehensive list of all vascular

plant species growing at a particular wetland at the

time of sampling and to characterize the relative

dominance of these species at several levels of scale

(basically herbaceous, shrub, small tree, and large

tree scales, or at 1 m2

, 10 m2, 100 m2, and 0.1 ha

(1,000 m2 or 1 are).

All vascular species within the modules are identified to species. Immature plants or plants missing

structures (e.g., fruiting bodies, etc.) that cannot be

identified to species are identified to genus or family or noted as unknown. Within the intensively

sampled modules, percent cover is recorded for

each species within modules and nested quadrats.

Cover classes suggested by Peet et al. (1998) are

used as a faster and more repeatable method for

assigning cover values: Class 1 (solitary/few), Class

2 (0 to 1%), Class 3 (1% to 2%), Class 4 (2%-

5%), Class 5 (5%-10%), Class 6 (10%-25%),

Class 7 (25%-50%), Class 8 (50%-75%), Class 9

(75%-95%), Class 10 (95%-99%). The midpoints

of the cover classes are used to calculate percent

cover, relative cover, etc.

Woody stem data (trees, shrubs, and woody lianas reaching breast height or 1.4 m) are collected

as counts of individuals in diameter classes. Peet

et al. (1998) suggest the following diameter classes

(in cm): 0-1, 1-2.5, 2.5-5, 5-10, 10-15, 15-20,

20-25, 25-30, 30-35, and 35-40, with stems

greater than 40 cm counted individually and measured to the nearest centimeter. Multiple stems arising from a common root system are recorded separately if they branch below 0.5 m from the ground.

Peet et al. (1998) recommend that the area surveyed by stem count be adjusted based on conditions at the site, e.g., reduced to 20% of the modules for dense shrubland or increased by 200% for

oak savannahs. This is easily implemented by reducing the width of the modules for woody species

only.

An important part of vegetation surveys is collection, preparation, and depositing of voucher specimens in major herbariums in order to document a

permanent record of that plant at that location. Although staff resources make collecting vouchers of

every vascular plant infeasible, a voucher specimen

of at least 10% of the vascular plant species at any

given site is collected; however, in every instance in

which the identity of any species cannot be confirmed in the field, or where field personnel disagree
70

as to the identity of a species, a voucher specimen

is collected for identification in the office. In particular, difficult genuses and families (e.g.,

Cyperaceae, Poaceae, Ranunculaceae, Viola, Aster, Potamogeton), as well as endangered, threatened, rare, or otherwise unusual plants, are almost

always collected for confirmation.

Finally, data on standing biomass for emergent

wetlands are collected. These data can be used in

several ways. Biomass production in emergent

wetlands dominated by herbaceous vegetation is

estimated by harvesting 900-cm2

 quadrats in each

wetland. The quadrats are located within the intensive modules of each plot. The plants within each

quadrat are cut at the soil surface and placed into

paper bags. In the lab, plants are oven dried at

105o

C for at least 24 hours and then weighed.

Lessons Learned for

Vascular Plants

Floristic Quality Assessment Indexes

Ohio EPA has found that the FQAI score and

subscores of the FQAI, e.g., percent coverage of

plants with coefficients of conservatism of 0, 1,

or 2, are very successful attributes and metrics

for detecting disturbance in wetlands. See the following references:

Andreas B, Lichvar R. 1995. A Floristic Quality

Assessment System for Northern Ohio. Wetlands

Research Program Technical Report WRP-DE-8.

U.S. Army Corps of Engineers, Waterways Experiment Station.

Herman KD, Masters LA, Penskar MR, Reznicek

AA, Wilhelm GS, Brodowicz WW. 1996. Floristic Quality Assessment With Wetland Categories

and Computer Application Programs for the State

of Michigan. Michigan Department of Natural Resources, Wildlife Division, Natural Heritage Program.

Wilhelm GS, Ladd D. 1988. Natural area assessment in the Chicago region. Transactions 53rd

North American Wildlife and Natural Resources

Conference, pp. 361- 375.

Wilhelm GS, Masters LA. 1995. Floristic Quality Assessment in the Chicago Region and Application Computer Programs. Lisle, IL: Morton Arboretum.

Semiquantitative disturbance/integrity scales

Ohio EPA has had good success in developing a

semiquantitative disturbance/biological integrity scale

called the Ohio Rapid Assessment Method for

Wetlands v. 5.0. Until more quantitative variables

such as percent impervious surface are found, this

type of tool is a good candidate for the problematic

x-axis in wetland biocriteria development. See also

“Plants and Aquatic Invertebrates as Indicators of

Wetland Biological Integrity in Waquoit Bay Watershed, Cape Cod,” Carlisle BK, Hicks AL, Smith

JP, Garcia SR, Largay BG. Environ Cape Cod

1999; 2(2):30-60, where a similar system was used

to rank levels of disturbance.

Classification

Classification is definitely an iterative process.

Investigators should consider a hydrogeomorphic

(HGM) classification scheme if one has been developed for their region of interest, at least as a starting point. However, the experience in Ohio suggests that grosser classes based on dominant vegetation (emergent, scrub-shrub, forested, etc.) may

work also. A goal of a cost-effective biocriteria

program is to have the fewest classes that provide

the most cost-effective feedback. With vegetation,

data from Ohio are suggesting that somewhat diverse wetland types may be “clumpable,” because

even though their flora are different at the species

level, the quality/responsiveness of their unique flora

to human disturbance is equivalent. This is also a

concern in States with high degrees of wetland loss

where too few wetlands of a particular HGM class

remain to analyze as a separate class.
71

Field and lab methods

After experimenting with both transect/quadrat and

releve-style plot methods, Ohio has adopted a plotbased method that allows for a qualitative stratification of wetland by dominant vegetation communities. This method is flexible and adaptable to

unique site conditions, provides dominance data for

all species in all strata, provides data that are

intercomparable with other common methods, is

relatively easy to learn, and is relatively fast and

cost-effective (up to two to three plots can be completed in a day).

Whatever sampling method is adopted, it is essential that dominance and density information

(cover, basal area of trees, stems per unit area, relative cover, relative density, importance values, etc.)

be collected. Many of the most successful attributes

Ohio has found in developing a vegetation IBI are

based on cover data of the herb and shrub layers

and density data of the shrub and tree layers.

Definitely consider using cover classes in general

and a class scheme that works on a doubling principle to aid in consistent inter-investigator usage (see

Peet et al. 1998). Then use the midpoints of the

class for your analysis. This seems to help with

consistent usage and smoothing out the roughness

in cover data.

Finally, it is recommended that initially the

sampling method should “overstratify” in both the

vertical and horizontal dimensions until it can be determined which strata and communities are responding best to human disturbance. Ohio has found that

the herb and shrub (subcanopy layers) seem to

respond the best, although some intermediate tree

size classes (e.g. 10- to 25-cm dbh) also appear

responsive.

Overstratifying horizontally may also make sense

at the reference development stage; however, ultimately the decision whether to split or clump communities depends on whether this is necessary to

detect the disturbances. “Homogenizing” community types by placing a releve or transect across

them (e.g., aquatic bed to emergent to shrub zone)

can be appropriate if splitting does not matter to

detect the disturbance. The caveat, of course, is

that you cannot separate the data set later if you

detect something of interest in one of the clumped

communities.

Vouchers and QAQC

On the basis of Ohio’s experience, voucher as

much as you can for later confirmation in the lab

and deposit vouchers in local and regional

herbariums. Definitely collect all Cyperaceae,

Poaceae, and Juncaceae, and also consider collecting shrubs genuses and families (Salix, Viburnum, Vaccinium, Rosa, Alder, etc.), Polygonum

spp., Aster spp., Viola spp., and Cryptograms.
72

Oregon: Simultaneous Development,

Calibration, and Testing of HydrogeomorphicBased (HGM) Assessment Procedures and

Biological Assessment Procedures

Purpose

 Collect and analyze field data on wetland and

riparian plant communities and hydrogeomorphic (HGM) features to define reference

standard conditions and performance criteria

for four subclasses of wetlands in Oregon, two

in the Willamette Valley in western Oregon and

two on the Oregon coast

 Use that information to develop quantitative but

rapid, HGM-based assessment models and

procedures for those subclasses, as well as to

identify new wetland plant community metrics

that have demonstrated relationships with landscape condition and with internal alterations of

wetlands in this region

 Compare results of using HGM models for functions with metrics that describe floristic characteristics, to verify the results on independent data

sets, and demonstrate implementation of the

methods for assessing progress of restored

wetlands and for helping recommend compensation ratios in a wetland mitigation bank context

Contact Dana Field

Organization Oregon Division of State Lands

775 Summer St. NE

Salem, OR 97310

Phone (541) 378-3805, ext. 238

E-mail Dana.Field@dsl.state.or.us

Contact Paul Adamus, Ph.D

Organization Adamus Resource Assessment,

Inc.

6028 NW Burgundy Drive

Corvallis, OR 97330

Phone (541) 745-7092

E-mail adamus7@attbi.com

Wetland Type

 Slope, flats, and estuarine fringe wetlands

Assemblages

 Vascular plants

Status

Completed reports for the two Willamette Valley

subclasses, and currently initiating development

of the estuarine fringe project. Information on

purchasing the reports can be found at:

http://statelands.dsl.state.or.us/hgm_

guidebook.htm.

Project Description

These projects are being done cooperatively with

EPA Region 10 and the Oregon Division of State

Lands, which in 1997 identified a need, in the context of Section 404 and 401 responsibilities, for a

more quantitative assessment method tailored specifically to these wetland subclasses. The proce-
73

dures have been designed to be applicable to

nonjurisdictional riparian sites as well as to wetlands. The project began in 1998 with formation of

an interagency oversight committee, intensive review of regional literature, and reconnaissance and

preliminary selection of 124 reference sites throughout the Willamette Valley ecoregion. In 1999, the

list of sites was narrowed, workshops of local experts were held to develop function models and field

procedures, 38+ volunteers were recruited and

trained to assist with field work, and 2 assessment

teams (“A-teams”) of scientists with help from the

volunteers collected data from the selected sites. A

three-volume final report was published in early

2001 (see above). The report for the estuarine

fringe wetlands is scheduled for completion in 2004.

Under administrative rules recently reissued by the

Oregon Division of State Lands, applicants for most

State-authorized wetland removal-fill permits are

now required to use the regionalized HGM classification that was developed by this project, and are

encouraged to use the reference-based function

assessment procedures when available for a particular subclass and region of the State.

Study Design

For the Willamette Valley project, plant and

hydrogeomorphic data were collected from 109

wetland and riparian sites (54 riverine impounding

subclass, 55 slope/flats subclass). The “riverine

impounding” sites include all wetland and riparian

areas within a 2-year river floodplain, e.g., sloughs,

oxbows, cut-off channels, beaver impoundments,

stream-fed ponds with water control structures.

“Slope/Flats” sites include most ash swales, wet

prairies, springs, and foothill seeps. From visual

inspection alone, most sites in the “slope” HGM

class could not be reliably separated from most sites

in the “flats” HGM class, due to the flat topography

and varied geology of the region, so the two were

combined into a “slope/flats” subclass.

Within the subclasses, assessment sites were selected nonrandomly, in a manner intended to span

gradients of human disturbance, size, and plant community succession. Restored/created sites of various ages were also included. Nearly all sites were

on public lands, and ranged in size from 0.1 to 233

acres. Most sites were nominated by local wetland

experts. From the collected data, five least-disturbed riverine sites were selected as reference standards, as were six least-disturbed slope/flat sites.

Use of cluster analysis and other statistical analysis

methods verified that sites belonging to the two main

subclasses were significantly different based on landscape position and mapped soil characteristics important to ecosystem function.

For defining the disturbance gradient (the “x axis”),

information specific to each candidate site was collected during the reconnaissance phase. This information pertained to past management practices,

surrounding land use, and recent physical alterations.

Physical alterations were noted visually during reconnaissance visits. At each site each type of alteration was categorized as (a) absent, (b) physically

affects <10% of site, (c) affects >10% of site but

not all, or (d) affects entire site. The categories that

were used for alterations were:

 Flow-impounding – berms, dikes, dams

 Flow-impounding – excavations, pits

 Flow-redirecting

 Water subsidy (e.g., stormwater pipes)

 Drainage-inducing (ditches, tile)

 Soil compacting (e.g., fill, machinery, cows)

 Soil mixing (e.g., plowing)

 Soil grading (e.g., flattening)

 Vegetation removal (e.g., extreme grazing, logging)

Sampling Methods: Plants

At each site, the A-teams identified plants in each

of potentially three hydrologic zones: permanent

water zone, seasonally inundated zone, saturated-
74

only zone. For woody plants, the team walked the

entire site and made an overall estimate of relative

percent of the area of each zone occupied by each

shrub species (understory and open) and tree species. For herbaceous plants, the team assessed

relative cover of each species in 1x1-m quadrats.

No more than nine quadrats were used at any site

(and fewer at smaller sites and sites with fewer hydrologic zones). Larger numbers of quadrats were

not used because of time and resource constraints.

Within zones, quadrats were located so as to maximize the cumulative number of species found. This

less formal approach was used because our goal

was to develop metrics based mainly on community composition rather than quantitative measures

of abundance or cover. Two teams assessed all

109 sites in about 50 workdays.

Analytical Methods: Plants

From the collected data, the following plant metrics

were compiled for each site:

 Number of native herb species, relative to the

intensity of sampling (# of plots at each site)

(based on analysis using species-area curves

and regression)

 Percent of herb species that are native

 Percent of the herb species that are “remnantdependent” (i.e., reputedly most characteristic

of unaltered sites)

 Percent of the dominant herb species in any plot

that are natives

 Percent of the dominant herb species in any plot

that are remnant-dependent

 Percent of the true wetland species (facultative

or wetter) that are natives

 Percent of shrub species that are natives (when

shrubs present)

 Percent of native shrub species that are at least

moderately dominant

 Percent cover of non-native shrubs

Preliminary analysis suggests many of the above

metrics had a statistically significant association with

categorical observations of partial physical degradation of sites (see Study Design, above) and/or

with amount and proximity of surrounding land cover

categories (agriculture, urban, natural) that were

assessed visually during fieldwork as well as by a

GIS analysis of existing digital imagery. Analyses

of the HGM and plant data sets, each containing

4,000+ records, were performed on a PC using

Excel, PC-ORD, and NCSS.

Lessons Learned

 Random or systematic sampling, whether within

a region or within a site, is not always appropriate for use in identifying good biological indicators or developing rapid models for assessing wetland condition and function.

 Systematic, repeatable, rapid procedures can

be developed for assessing some of the disturbance gradients. This is a necessary precursor

to selecting reference sites that will yield the most

useful data.

 The biological metrics investigated or used

should be appropriate to the study design and

measurement protocols.

 Data suitable for identifying biological indicators of wetland condition sometimes can be collected simultaneously with data collected for

calibration of HGM models. This does not necessarily require a great deal of additional training or field time.

Shared field experiences are a good forum for

sharing wetland knowledge among agencies, and

among agencies and consultants and citizens.

Shared field experiences lead to participants feeling more vested in the process of developing models and multimetric indexes. This informal “buy-in”

can lead to greater willingness of participants to use

the methods that ultimately result.
75

Pennsylvania: Assessing Wetland Condition at

a Watershed Scale Using Hydrogeomorphic

Models and Measures of Biological Integrity

Purpose

 Develop biological and functional assessment

methodologies for many wetland types in Pennsylvania

Wetland Type

 Variety of wetland types

Assemblages

 Amphibians

 Birds

 Macroinvertebrates

 Vascular plants

Project Description

The Penn State Cooperative Wetlands Center

(CWC) has numerous ongoing projects that involve

bioassessment methodology for assessing the condition of wetlands. These projects include:

 Bird community index of biological integrity for

the Mid-Atlantic Highlands

 Verified suitability index for the Louisiana waterthrush

 Amphibian indicator landscape study

 Macroinvertebrates indicator pilot study

 Otter and beaver interactions in the Delaware

Water Gap

 Using bioindicators to develop a calibrated index of regional ecological integrity for forested

headwater systems

In addition to these projects, CWC is conducting

a 2-year pilot study to assess the ecological condition of wetlands in the Juniata River watershed of

central Pennsylvania. The Juniata River is a major

tributary of the Susquehanna River and lies within

the headwaters region of the Chesapeake Bay ecosystem. The objective of the Juniata Wetland Monitoring Project is to define the ecological health of

wetland resources, thereby providing a scientific

context for resource managers to plan future protection and restoration activities.

Contact Denice Heller-Wardrop

Organization Penn State University

Penn State Cooperative Wetlands

Center

301 Forest Resources

Laboratory

University Park, PA 16802

Phone (814) 863-1005

E-mail dhw110@psu.edu

Website http://www.wetlands.cas.psu.edu/

Contact Rob Brooks

Organization Penn State University

Penn State Cooperative Wetlands

Center

301 Forest Resources

Laboratory

University Park, PA 16802

Phone (814) 863-1596

E-mail rpb2@psu.edu

Website http://www.wetlands.cas.psu.edu/
76

CWC is using HGM functional assessment models that incorporate a floristic quality assessment

index (FQAI) similar to the one used by Ohio EPA

(see Ohio EPA’s case study). They use a threestep process. The first step is to develop and calibrate HGM functional assessment models that are

sensitive to environmental disturbance. The second step is to randomly sample wetlands within the

watershed that represent a range of HGM classes

and disturbance levels. The final step is to apply the

HGM models to the data collected in the field to

determine the condition or health of wetlands within

the Juniata River watershed.

Preliminary HGM functional assessment models

have been developed by the CWC. These models

are currently being calibrated using a set of 102

reference wetlands that span a variety of HGM subclasses (mainstem floodplain, headwater floodplain,

riparian depression, slope, etc.) and disturbance

levels (severe, moderate, or pristine). Disturbance

levels are assigned based on the surrounding landscape, the type of buffer present, and potential stressors identified at the site. It is essential to characterize wetlands across a disturbance gradient to

determine not only the level of function a given wetland type may achieve, but also the level of functioning that is attainable in an impacted landscape.

The second step of the process, data collection,

is being undertaken by the CWC and trained interns from Penn State University, the University of

Pittsburgh at Johnstown, and the Department of

Environmental Protection. Two types of data are

collected for each wetland sampled: landscapelevel data and site-level data. Landscape-level data

are obtained at the CWC by characterizing land

uses within a 1-km radius circle surrounding the

wetland using aerial photographs. This gives an indication of the type and magnitude of potential wetland stressors. For example, activities such as land

clearing in the surrounding watershed may stress

wetland systems by increasing sediment loads.

In addition to land uses, the buffer type and width

is also determined for each site. It is important to

look at buffers when studying disturbance because

buffers can ameliorate or magnify the affects of negative landscape activities. A wetland surrounded by

a forested buffer may not be as stressed by activities in the landscape as one surrounded by an urban or agricultural buffer.

Site-specific data are collected at each wetland

site in the field. Site-level responses of wetlands to

stressors are many and may involve both abiotic

and biotic indicators. At each wetland sampled, data

are gathered on microtopography, soils, plants, and

animals. Although not all plant species are highly

sensitive to disturbance, the immobility of the plant

community, its amenity to remote sensing techniques, and easily recognized signs of stress make

it a good indicator of wetland condition. Previous

studies by the CWC have investigated the potential

utility of plant community measures as indicators of

wetland health. Plant community measures that may

prove to be good indicators of disturbance include

the FQAI score, the number or dominance of introduced or aggressive native species, and the number of different species of Carex present. The CWC

method contains a comprehensive plant sampling

methodology that has been used in a variety of

project types. Three sizes of plots are used to

record various measures of the plant community

including percent cover and richness of herbaceous

species, shrub volume, and tree dbh.
77

Vermont: Classification, Biological

Characterization, and Biometric Development

for Northern White Cedar Swamps

and Vernal Pools

Contact Doug Burnham

Organization Vermont Department of Environmental Conservation

WQD

103 S. Main St. – 10N

Waterbury, VT 05676

Phone (802) 241-3784 or 244-4520

E-mail DOUGB@dec.anr.state.vt.us

Purpose

The Vermont Wetlands Bioassessment Project is

a collaboration between the Vermont Department

of Environmental Conservation and the Vermont

Nongame and Natural Heritage Program, with a 3-

year time frame. The primary objectives of the first

phase of the Vermont Wetlands Bioassessment Program are:

 Gather chemical, physical, and biological data

from seasonal pools that will facilitate an ecologically based classification of minimally disturbed (reference) seasonal pools in Vermont

 Use both previously and newly collected Nongame and Natural Heritage Program data to

try to identify specific biological attributes that

can serve as indicators of ecological integrity in

northern white cedar swamps.

Wetland Type

 Vernal pools

 White cedar swamps

Assemblages

 Algae

 Amphibians

 Birds

 Macroinvertebrates

 Vascular plants

Status

Analyzing data and writing final report. A final

report is expected in the near future.

Project Description

The Vermont Wetlands Bioassessment project currently focuses on two types of wetlands: (1) vernal

(seasonal/ephemeral) pools and (2) Atlantic northern white cedar swamps.

Vernal pools

We sampled a total of 28 vernal pools for breeding amphibians, macroinvertebrates, algae (diatoms),

soils, vegetation, and water chemistry in April, May,

and June of 1999 and 2000. A variety of other

physical and riparian observations were also made.
78

Pools were distributed throughout Vermont across

seven biophysical regions. A range of condition,

from “reference” or least disturbed, to highly disturbed, was represented in the selection of pools.

Upon completion of the field phase of the project

and assessment of relative hydroperiod, it is likely

that 5 of the 28 pools may be less “seasonal” than

originally thought. Data are being analyzed for biological signals that may identify pools with moreor-less permanent standing water. Each pool was

visited twice, the two visits approximately 4 to 6

weeks apart, for aquatic macroinvertebrate and

water chemistry sampling. Algae samples were collected on the second pool visit. Amphibian surveys

either preceded or coincided with the first

macroinvertebrate sampling visit. Five of the 28

pools were dry on the second sampling visit.

Aquatic macroinvertebrates. Macroinvertebrates were sampled using three different methods:

funnel traps to sample the actively swimming invertebrates (i.e., beetles, bugs, mosquitoes, crustaceans), a D-net to sample benthic invertebrates in

the leaf litter and muck (i.e., snails, bivalves, chironomids, oligochaetes, caddisflies), and a qualitative search for any taxa we might have missed with

the previous two methods. Funnel traps were made

of window screen and designed to function like minnow traps. The traps were placed approximately

10 m apart and were left in place for approximately

24 hours. When the traps were emptied, any amphibians were returned to the pool and the

macroinvertebrates were collected and preserved.

The contents of each trap were stored separately.

The D-net scoop and qualitative samples were preserved in the field, and later picked and sorted into

taxonomic orders according to standard protocol.

Except for zooplankton, all taxa were identified to

the lowest practicable level. Scoop sampling along

transects often created more of a disturbance in the

pool than was comfortable for those conducting the

sampling. The presence of egg masses and/or large

concentrations of larval amphibians greatly restricted

the efficiency of the scoop sampling at many pools.

Trap sampling was less disruptive, but care had to

be taken to not completely submerge the traps when

adult amphibians were present so that entrapped

adults would have air space to breathe. High concentrations of mosquito larvae tended to affect the

efficiency of the traps. Qualitative sampling was

less disruptive (although excessive wading in the

pools often created an uncomfortable level of disturbance) but provided an inconsistent sampling effort. The majority (70%) of macroinvertebrate taxa

collected were represented in trap samples. Two

orders represented a large proportion of all taxa

collected: Diptera (35%) and Coleoptera (23%).

Water. Water temperature, pH, and apparent

color were recorded in the field. The remainder of

the water sample was preserved and later analyzed

for alkalinity, conductivity, anions, cations, and aluminum, and again for pH. Field pH values ranged

from a minimum of 4.41 to a maximum of 7.75,

alkalinity values ranged from 0 mg/L to 173.0 mg/

L, conductivity readings ranged from 14.9 umhos/

cm to 335 umhos/cm, and apparent color ranged

from 4.5 to 489 over the sampling season. Apparent color, alkalinity, and cation concentrations show

a general increasing trend within each site over the

sampling season, whereas pH and aluminum and

anion concentrations exhibited no consistent trend.

Algae. Algae were collected and analyzed for

diatoms from 18 pools sampled in 1999. Algae sampling primarily targeted diatoms; however, filamentous algae were collected when present. We attempted to collect both benthic samples (scraping

algae from leaves, sticks, and rocks) and planktonic diatom samples from each pool. Unfortunately, five of the pools were already dry in May,

so we were only able to collect planktonic samples

in the remaining 13 pools. Benthic samples were

collected from 17 of the 18 pools. We froze all

samples upon return to the lab, and samples were

sent to Dr. Jan Stephenson for identification.

Amphibians. We observed amphibians initially

during the amphibian survey, and continued to observe and collect specimens during both rounds of
79

macroinvertebrate sampling. At the beginning of

the field season, we visually surveyed each pool for

egg masses and spermatophores, identified each egg

mass, recorded an approximate number of eggs per

mass, counted and identified breeding adults, and

described physical parameters of the habitat. The

timing of this first visit did not necessarily coincide

with amphibian emergence; however, the funnel traps

used during invertebrate sampling effectively caught

active amphibians. Breeding adults were typically

captured during the first round of sampling, whereas

tadpoles and larvae were present in the traps on

the second visit. All amphibians caught by the funnel traps, including adults, larvae, and egg masses,

were identified in the field, counted, and returned

to the pool. We continued to survey egg masses

and record physical parameters throughout the field

season. All 28 pools sampled showed signs of use

by breeding amphibians. Amphibian species commonly observed included wood frogs (27/28 sites),

yellow-spotted salamanders (27/28 sites), Jefferson

(hybrid) salamanders (7/28 sites), red-spotted newts

(18/28 sites), and green frogs 15/28 sites).

Ongoing and upcoming work. Data are currently being evaluated using a variety of procedures

in order to identify and describe efficient and least

disruptive sampling methods, and evaluate biological, chemical, and physical indicators of natural variability and disturbance. Methods of data analysis

include two-way indicator species analysis

(TWINSPAN) and detrended correspondence

analysis (DCA) as well as standard descriptive and

comparative statistical methods. A detailed site report is being developed for each vernal pool site.

Northern white cedar swamps

Seven northern white cedar swamps were surveyed in June for breeding birds and vegetation.

Two of these cedar swamps were considered to be

of reference quality, whereas the other five had some

degree of impairment associated with them. Disturbances at the impaired sites included logging,

roads, and storm water and agricultural discharge.

We visited three of the sites early in summer to

assess the feasibility of sampling aquatic

macroinvertebrates for bioassessment and monitoring purposes.

Breeding birds/vegetation. Vegetation and biophysical data were collected at each site and species lists are being constructed. Listening stations

were established and a bird census was taken twice

during the breeding season. The data collected from

the vegetative, biophysical, and bird sampling will

be compiled and compared to the existing data from

the statewide inventory of northern white cedar

swamps.

Aquatic macroinvertebrates. Sampling for

aquatic macroinvertebrates had been scheduled for

May, but because of a busy vernal pool sampling

season, we were unable to visit the cedar swamps

until June. Unfortunately, a very dry spring may

have caused the cedar swamps to be drier than usual

this year. We visited two impaired sites and one

reference site during the month of June. We were

unable to find any standing or flowing water at either the reference site or at one of the impaired sites,

so we could not effectively sample. However, we

found evidence that suggested the swamps had contained water earlier in the season. The second impaired site contained many braided, slow-flowing

channels and some standing water. We qualitatively

sampled three of the channels and a small hollow at

the base of the boulder. The samples were preserved in the field, picked, and sorted, and will be

identified according to standard protocol. It is possible that aquatic microhabitats in cedar swamps

are not available consistently enough to sample for

aquatic macroinvertebrates; however, our limited

sampling efforts did not conclusively elucidate the

feasibility of using aquatic macroinvertebrates as

biological indicators in cedar swamps.

Ongoing and upcoming work. Using the data

we have collected, we will work to identify attributes

associated with vegetation and bird assemblages

that can serve as indicators of ecological integrity

and anthropogenic disturbance.
80

Washington: King County Wetland-Breeding

Amphibian Monitoring Program

Contact Klaus O. Richter

Organization King County Department of Natural

Resources & Parks

Water and Land Resources Division

201 S Jackson Street, Suite 600

Seattle, WA 98104-3855

Phone (206) 205-5622

E-mail klaus.richter@metrokc.gov

Organization Viewpoint Services

11040 104th Ave. NE

Kirkland, WA 98033-4423

Phone 425-822-4016

E-mail korichter @msn.com

Purpose

Provide yearly data regarding wetland condition

and the health of breeding amphibians for environmental review, to identify threats to wetlands and

amphibians, to evaluate current wetland protection

measures, and to improve local-land use decisions.

Specifically, to stem amphibian extinctions in urban

landscapes by first identifying cause-and-effect relationships among amphibian distribution, abundance, and health and anthropogenic impacts to

wetlands and uplands, and second, using this information to provide amphibian conservation recommendations.

Wetland Type

 Depressional wetlands

Assemblages

 Amphibians

Status

 Implementing methods

Project Description

The King County Wetland-Breeding Amphibian

Monitoring Program was originally designed to provide the county with long-term wetland, amphibian,

and landscape information for planning and regulatory purposes. From 1993 to the present, a minimum of 150 volunteers were trained to census amphibian eggs, juveniles, and adults in 90 freshwater

wetlands of 26 watersheds in King County. Analysis of amphibian distribution and health for all wetlands was completed. A targeted subsample of 21

wetlands in 3 rapidly urbanizing watersheds in which

amphibians were declining was additionally monitored for wetland hydrology, predators, and watershed condition to determine the causes of population declines, in hope of stemming declines through

better wetland conservation practices.

Today, a skeleton program totally run by volunteers continues to monitor a few of the initially surveyed wetlands without county support. A more

intensive monitoring program targeting amphibians

as bioindicators of wetland condition has been initiated with the cooperation and partnership of the

University of Washington’s Certificate Program in
81

Wetland Science and Management, in which several students research and monitor an important

wetland and amphibian issue.

The project’s monitoring goals include the

following:

 Identify the occurrence of the State-endangered

Oregon spotted frog

 Determine land uses compatible with wetland

and amphibian conservation objectives

 Provide data to help develop and implement

regulations for the protection of amphibians and

their habitats

 Identify population distribution status of other

county declining species

 Obtain standardized baseline inventory data on

the distribution, abundance, and health of amphibians in King County wetlands

 Provide information to King County, Washington State Department of Fish and Wildlife,

Washington State Department of Ecology, and

Federal resource agencies for developing regional wetland and wildlife management programs

 Develop an effective public outreach and education program to train citizens to monitor amphibians and wetland conditions to foster wetland stewardship

 Educate potential wetland scientists to the diversity of wetland issues and train them in developing monitoring programs, assessing wetland condition, and crafting management plans

for better wetland conservation

 Investigate local urban amphibian decline and

extinction issues

 Develop methods for utilizing amphibians as

bioindicators of wetland condition by establishing cause-and-effect relationships between

amphibian declines and wetland hydrology,

water quality, predators and pathogens, and

watershed land use.
82

Wisconsin: Developing Biological Indexes for

Wisconsin’s Palustrine Wetlands

Contact Jennifer Hauxwell

Organization Wisconsin Department of Natural Resources

Bureau of Integrated Science Services

1350 Femrite Drive

Monona, WI 53716

Phone (608) 221-6338

E-mail hauxw@dnr.state.wi.us

Purpose

 Develop a biological index for Wisconsin’s

palustrine wetlands

 Compare performance of one plant and two

macroinvertebrate multimetric indices

 Develop biological integrity rating system for

classifying wetlands

Wetland Type

 Palustrine wetlands

Assemblages

 Macroinvertebrates

 Vascular plants

Status

Findings representing preliminary investigations are

presented in Final Report to EPA Region V, Wetland Grant #CD985491-01-0 prepared by Wisconsin DNR (January 2000).

Project Description

Field studies for this project were conducted during the spring and summer of 1998, with laboratory

analysis and data synthesis completed the following

year. Funding was provided by a grant from EPA

Region 5. The findings formed the basis for a second EPA-funded grant to refine and further evaluate the preliminary indices and expand communities covered to include amphibians, small mammals,

diatoms, and zooplankton.

Study Design

We sampled 104 palustrine depression wetlands

distributed across the major ecoregions of Wisconsin

during early spring and summer of 1998. Study

sites included a mixture of least-disturbed reference

basins and degraded or restored wetlands, representing a range in vegetative cover types, water

chemistries, and water duration.

Sampling Methods:

Macroinvertebrates

We sampled macroinvertebrates early in the spring

to minimize influences of immigration-emigration.

Three different field collection procedures were
83

evaluated and two laboratory approaches were

used. On all 104 wetlands, we collected two sets

of three standard D-frame net sweeps of approximately 1-m length each. Sampling stations (< 60

cm deep) were established at equally spaced points

around the wetland perimeter that approximately

trisected the basin (and ensured coverage of the

major plant communities present). The first set of

net sweeps was concentrated (large coarse materials were rinsed, examined, and removed) into a 1-

quart container; the second set of net sweeps was

not field-concentrated, but rather was placed directly into a 1-gallon plastic bag. Both sets were

preserved in ethanol and returned to the laboratory

for processing. The A third set (used on a subset of

17 wetlands) of D-frame net sweep samples was

placed on a coarse wire screen over a collection

basin for a period of approximately 10 minutes.

Organisms falling (or crawling) through the screen

and entering the collection container were collected

and preserved as above. This last set represented

a “clean” sample that was much easier to sort and

process than the standard samples.

Laboratory Methods:

Macroinvertebrates

Macroinvertebrates were processed using a twotiered approach. The first stage consisted of a fixed

100-count (sensu Hilsenhoff Biotic Index procedures) using a grid-marked tray with 24 cells. Organisms were picked and sorted at a coarse taxonomic level, usually order or family only. Following

completion of the 100-count sample, we processed

the balance of the sample in its entirety (except for

subsampling dominant taxa). The unconcentrated

“bag” samples proved to be too large to process in

an economical fashion, so only the complete set of

104 “field-concentrated” samples were processed.

The 17 “screened” samples generally contained less

than 100 total organisms and were processed completely.

Analytical Methods:

Macroinvertebrates

We used SYSTAT to perform all statistical and

graphical analyses. Percentage data were transformed using the arc-sine square-root transformation, and abundance data were either log-transformed or power-transformed as applicable. Metric development was based on a series of visual

comparisons of community attribute responses to

suspected measures of disturbance using box plots

and jittered dot density plots. Those attributes that

exhibited evidence of separation between reference

wetland conditions and wetlands suspected to be

impacted by human disturbance were selected as

potential metrics. Attributes that exhibited inconsistent or overlapping responses between impacted

and reference systems were eliminated from further

consideration.

Sampling Methods: Plants

We conducted simplified plant surveys during July

1998 using a combination of techniques. This included a subjective estimate of cover and an objective survey of percent cover and frequency of

occurrence within six equidistantly spaced 20 by

50 cm rectangular quadrats positioned along each

of three transects that trisected the wetland basin

(total of 18 quadrats per wetland).

Laboratory Methods: Plants

No biomass or stem counts were performed.

Voucher specimens were pressed and identified to

species when possible, but most plant metrics were

based on a coarser taxonomic level.
84

Analytical Methods: Plants

We developed the plant biological index using the

same procedures described for the two

macroinvertebrate-based indices. Because we

wanted to develop a practical tool for managers

with limited botanical expertise, we lumped taxa at

various taxonomic levels (e.g., family, genus) or

structural groups (e.g., grass-like, emergent) for

analysis. Importance values (average of percent

cover and frequency of occurrence) were used as

the attribute of concern for family-genus-species

levels, and percent cover was used for emergent,

submergent, floating-leafed, and open-water

attributes.

Other Data Collected

We also collected associated physical and chemical data on each wetland, including pH, alkalinity,

conductivity, color, temperature, clarity, and depth.

Riparian cover type within a 100-foot buffer area

surrounding each wetland was subjectively estimated

and recorded, as was shade canopy cover.

Preliminary Findings

Three multimetric indices (two macroinvertebrate

and one plant index) were developed. The Wisconsin Wetland Macroinvertebrate Index (WWMI)

is a multimetric index based on 15 metrics derived

from a total count of organisms in three composited

net sweeps. A total of 69 community attributes were

evaluated. The WWMI is composed of 12 abundance metrics, two richness metrics, and one percentage metric. Abundance metrics include mollusks, annelids, fairy shrimp, damselflies, pigmy

backswimmers, water boatmen, limnephild

caddisflies, total caddisflies, phantom midges, mosquitoes, soldier flies, and total invertebrates. Richness metrics are noninsects and total taxa. The

percent caddisflies is the only percentage metric.

Apparent redundancies (e.g., caddisflies) in the

metric may or may not be an issue; differences in

taxonomic rate of development in wetlands due to

thermal dynamics may require a certain amount of

redundancy to ensure that important taxonomic

groups are accounted for. The WWMI is used to

rate, rank, or compare wetland biological condition.

The second macroinvertebrate index, termed the

100-count macroinvertebrate biotic index (100-

count MBI), is based on 10 metrics derived from a

random pick of 100 organisms found in the three

composited net sweeps. The 100-count MBI is

composed of 9 percentage metrics and 1 richness

metric (noninsect taxa). Percentage metrics include

pigmy backswimmers, water boatmen, total “bugs,”

limnephilid caddisflies, total caddisflies, chironomids,

soldier flies, and the sum of EphemeropteraOdonata-Trichoptera (EOT) taxa. Noninsects represent the only richness metric in the 100-count MBI.

The ninth percentage metric, mollusks, may be only

useful in prairie-type wetlands. The 100-count MBI

is intended to be applied in the field by experienced

staff as a means of rapid bioassessment.

The third index is the Wisconsin Wetland Plant

Biotic Index (WWPBI) is based on eight (or nine)

plant metrics derived from transect data (representing 18 quadrats) and is intended to serve as a supplementary index to the WWMBI to rate, rank, and

compare wetland biological condition. Of 24 plant

community attributes tested, only one richness

(count) metric, one percent metric, and seven importance value-based metrics demonstrated consistent and predictable response. The single richness metric, total taxa, may require further modification after reaching some consistency regarding

taxonomic resolution (currently mixed family-genusspecies). Importance value-based metrics included

Carex, reed canary grass, cattail, duckweed,

bluejoint grass, and “good” plants (the sum of a

group of plants including all Carex, Utricularia,

Potamogeton, Calamogrostis, Sagittaria,

Polygonum, and Equisetum). An additional importance value-based metric, “pondweeds,” would only

be applied to wetlands with water duration exceed-
85

ing 7 months per year. The only percentage-based

metric, floating-leafed plants, would likewise only

be applied to wetlands with water durations exceeding 7 months.

Lessons Learned

Macroinvertebrates

 Water duration is an important factor shaping

macroinvertebrate community composition and

derived metrics that must be accounted for in

metric scoring.

 A coarse level of taxonomic resolution (order

and family) appears to be satisfactory in developing wetland macroinvertebrate metrics.

 Issues relating to redundancy among metrics,

influences of water chemistry, differences among

ecoregions, and seasonal variations need to be

addressed in more detail. Undoubtedly, these

factors need to be accounted for in establishing

rating scores and/or in refining metrics for use

in different areas or habitats.

 Basic differences exist in macroinvertebrate

communities between wetlands representing

wooded kettle depressions and open-prairie

type depressions in Wisconsin.

 Our greatest difficulty was in selecting and assigning some measure of “human impact” to the

study site wetlands. Further research will be

required to quantify the degree of human impact in order to refine biological response

metrics and indices.

 The WWMBI was not stable across dates (and

not designed to be); consequently, its use is restricted to early spring.

 Each macroinvertebrate index has its own set

of advantages and disadvantages; further refinement is required to enable their successful

application in the field.

Plants

 Some biases were apparent in the WWPBI, as

reference kettles scored consistently higher than

prairie wetlands.

 WWPBI scores in restored prairie wetlands

were better than in many natural wetlands, suggesting that wetland restorations in Wisconsin

may be adequate in terms of “restoring” the

vegetative community (not true for

macroinvertebrate response).

 The WWPBI shows some promise in its performance and, because of its taxonomic simplicity, it could be applied by nonbotanists.

Followup: Continuing Work

With the assistance of a second EPA wetland grant,

we have refined the macroinvertebrate and plant

multimetric indices and explored expanding the community components to include zooplankton (in cooperation with Dr. Stanley Dodson, University of

Wisconsin–Madison), diatoms (Paul Garrison, Wisconsin DNR), amphibians, and small mammals.

Findings associated with the second grant are reported in the second Wisconsin case study below.
86

Wisconsin: Refinement and Expansion of the

Wisconsin Wetland Biological Index

for Assessment of Depressional

Palustrine Wetlands

Contact Jennifer Hauxwell

Organization Wisconsin Department of Natural Resources

Bureau of Integrated Science Services

1350 Femrite Drive

Monona, WI 53716

Phone (608)-221-6373

E-mail jennifer.hauxwell@dnr.state.wi.us

Purpose

 Test and refine a Biotic Index for Wisconsin’s

palustrine wetlands

 Expand list of assemblages to include

macroinvertebrates, zooplankton, diatoms,

amphibians, plants, and small mammals

 Establish a biological integrity rating system for

classifying wetlands based on the response of

selected biological attributes (metrics) of the

above communities to surrogate measures of

human disturbance

Wetland Type

 Palustrine wetlands

Assemblages

 Macroinvertebrates

 Aquatic plants

 Zooplankton

 Diatoms

 Amphibians

 Small mammals

Status

Findings presented in Final Report to EPA Region V prepared by Wisconsin DNR dated April

2002.

Project Description

This project represents the evaluation and expansion phase of an earlier study that resulted in the

preliminary development of a Wisconsin Wetland

Biological Index based on plant and

macroinvertebrate metrics (please see final report

to EPA -Wetland Grant #CD985491-01-0). Data

from this second study were used to refine and further evaluate the preliminary indices and expand

communities covered to include zooplankton, diatoms, amphibians, and small mammals. Field studies for this project were conducted during the spring

and summer of 2000, with laboratory analysis and

data synthesis completed in 2001. Funding was

provided by a grant from EPA Region 5.

We sampled 75 palustrine depression wetlands in

southeast Wisconsin during early spring and summer of 2000. Study sites included a mixture of leastdisturbed reference basins (17 prairie and 19
87

wooded kettles) and impacted wetlands (18 urban

and 20 agriculture), representing a range in vegetative cover types and water chemistries. A severe

drought, which began the previous year, caused most

of the smaller wetlands with short hydroperiods

(seasonal and temporary) to dry out, and consequently only long-duration wetlands (semipermanent and permanent) were sampled.

We sampled macroinvertebrates early in the spring

to minimize influences of immigration-emigration.

On each wetland, we collected three standard Dframe net sweeps of approximately 1-m length each.

Sampling stations (less than 60 cm deep) were established at equally spaced points around the wetland perimeter that approximately trisected the basin and assured coverage of the major plant communities present. The contents of the three net

sweeps were concentrated (large coarse materials

were rinsed, examined, and removed) into a 1-quart

container, preserved in ethanol, and returned to the

laboratory for processing. Seven field replicates

were collected.

Laboratory Methods:

Macroinvertebrates

Each macroinvertebrate sample was processed

entirely, using a two-phase approach with

subsampling employed only for extremely abundant

organisms. The first stage involved using a gridmarked tray with 24 cells. Cells were selected randomly, and organisms were picked and sorted at a

coarse taxonomic level, usually to order or family

level. The abundance of organisms present in the

first two or three randomly selected cells was used

to project the abundance of the most common taxa

in the sample. Taxa whose abundance was estimated to exceed 300 in the total sample were then

overlooked while the balance of the sample was

processed (the second phase of processing). All

specimens were vouchered (preserved in 70% ethanol) for possible further evaluation.

Sampling Methods: Plants

We conducted simplified plant surveys during July

2000 using a combination of techniques. This included a subjective estimate of cover and an objective survey of percent cover and frequency of

occurrence within six equidistantly spaced 20x50-

cm rectangular quadrats positioned along each of

three transects that trisected the wetland basin (total of 18 quadrats per wetland).

Laboratory Methods: Plants

No biomass or stem counts were performed.

Voucher specimens were pressed and identified to

species when possible, but most plant metrics are

based on a coarser taxonomic level.

Sampling Methods: Zooplankton

Note: zooplankton studies were conducted by Dr.

Stanley Dodson, University of Wisconsin-Madison,

Zoology Department, Madison, Wisconsin. E-mail:

sidodson@facstaff.wis.edu.

We collected one zooplankton sample from a central basin location in each wetland during June 2000

using a 5-L plastic bucket. We filtered a known

volume of water through a No. 10 (60-micron mesh)

net to capture zooplankton within. Seven field replicates were collected. Samples were preserved in

70% ethanol until processed.

Laboratory Methods:

Zooplankton

Each sample was scanned at moderate magnification for species of cladocera, copepods, ostracods, and aquatic insects. Slides and dissections

were made where necessary; for example, to aid in

the identification of copepod species. Total number of male and female Daphnia was counted in
88

each sample. More than 200 slides were prepared,

and identification of organisms is in progress. Organisms will be identified to species where possible.

Sampling Methods: Diatoms

Diatom studies were conducted under the direction of Paul Garrison, WDNR, Bureau of Integrated

Science Services, Environmental Contaminants

Section, 1350 Femrite Drive, Monona, WI 53716.

E-mail: garrip@dnr.state.wi.us

Using a 1-dram vial as a sample collection device, we collected and composited surficial (upper

0-1 cm) sediments from five sites in each wetland.

Seven field replicates were collected. Samples were

kept on ice or refrigerated until processed.

Laboratory Methods: Diatoms

Each diatom sample will be thoroughly mixed, and

a small amount will be placed into a tall beaker.

Hydrogen peroxide will be added and the sample

will be allowed to steep for about 5 minutes. Potassium dichromate will be added (under a ventilated hood and handled with safety gloves) to facilitate reduction of organic matter. The sample will

be washed at least four times with deionized water

by centrifuging for 10 minutes. Two portions of the

cleaned sample will be dried on separate No. 1

cover-slips and mounted with Naphraxâ, and labeled accordingly. Specimens from both cover slips

will be identified and counted under oil immersion

objective (1,400X) until a total of 250 frustules are

counted. Identification of difficult taxa will be made

using a scanning electron microscope. Undetermined specimens representing a significant portion

of a sample will be sent to Dr. Rex Lowe at Bowling Green University, Dr. Jan Stevenson at Louisville University, and/or Dr. Gene Stormer at the

University of Michigan.

Sampling Methods: Amphibians

Because amphibians are extremely sensitive to

weather and temperature, we assessed amphibian

communities during two separate sampling periods.

We conducted standardized frog-toad calling surveys (using WDNR protocols) during the first two

phenologies (early spring and late spring) between

the hours of 8 and 10:30 pm for 10-15 minutes

when water temperatures were above 50°F during

the first phenology or above 60°F during the second phenology. We recorded all calling to permit

verification of questionable identifications. In addition to calling, we added to the database any personal observations of amphibians made during any

of the daylight visits and any specimens captured

during the macroinvertebrate surveys.

Laboratory Methods: Amphibians

No laboratory methods were used.

Sampling Methods:

Small Mammals

Small mammal studies were conducted by R.

Bautz, WDNR, Bureau of Integrated Science Services, Ecological Inventory & Monitoring Section,

1350 Femrite Drive, Monona, WI 53716. E-mail:

bautzr@dnr.state.wi.us.

We assessed small mammal communities by trapping during August-September 2000. On each

wetland, we set 46 baited traps (mixture of 40 museum special grade and 6 larger tomahawk traps)

along transects (zigzag scattered routes) in the riparian zone (variable dimensions, depending on

setting) for one night. A one-day/night trapping

period was used to minimize disturbance by raccoons and other predators. Bait consisted of a mixture of peanut butter and rolled oats. Traps were

cleaned and rebaited each morning. Specimens

were placed in labeled freezer bags and returned to

the laboratory for identification.
89

Laboratory Methods:

Small Mammals

Identifications of rare taxa and species of special

concern were verified by local experts from the

University of Wisconsin.

Other Data Collected

We also collected associated physical and chemical data on each wetland. The water chemistry

measures included pH, alkalinity, conductivity, color,

chloride, calcium, silica, nitrate-nitrite, organic-nitrogen, and total phosphorus. Chemical analyses

were performed at the Wisconsin State Laboratory

of Hygiene following standard EPA-approved procedures. Physical data collected included water

depth, apparent color, temperature, and size. Riparian cover type within a 100-foot buffer area surrounding each wetland was subjectively estimated

and recorded, as well as shade canopy cover.

Analytical Methods: Data

Analysis and Development of

Biological Indices

Data were entered into Excel spreadsheet and

Access databases and examined for entry errors

(univariate checks) and anomalies (e.g., bivariate

plots) prior to analysis. Various community or species attributes (e.g., taxa or species richness, diversity, presence or absence of selected functional feeding guilds, trophic structure, percentages) were

evaluated and scored as potential metrics based on

their responsiveness to measures of human disturbance. Community attributes were examined using

a combination of procedures (outlined below) to

select promising metrics for index development.

Attributes that exhibited strong positive or negative

correlation with selected human response variables

were selected as metrics. We compared the sensitivity and correspondence among the selected community metrics, and developed a single multimetric

index of ecosystem integrity that related to overall

wetland condition.

Because different forms of human disturbance

elicit different responses among the various wetland biological communities, it is difficult if not impossible to choose one measure that represents “the”

single best measure of human disturbance. For example, the amphibian community may respond more

directly to woodland impacts (distance to nearest

woodland, patch size, corridor dimensions, etc.)

than to nutrient or pesticide inputs, whereas zooplankton and diatom communities may be more responsive to percent row crops in the watershed.

Consequently, we evaluated biotic responses to a

combination of a priori classes of human disturbance

and a composite chemical index that incorporated

nitrogen, phosphorus, and chloride concentrations.

The a priori classes represented agricultural and

urban impacts as well as subclasses of each, including three intensity levels of agricultural impact

and two forms of urban impact. Biotic responses

occurring within these classes were measured

against the responses in least disturbed kettles and

prairie depression wetlands. Statistical procedures

and approaches used in the analysis varied among

the communities assessed because of inherent differences in responses and the type of data collected.

Canonical correspondence analysis was used to

explore unimodal distributions of the diatoms along

the various environmental gradients. We used

SYSTAT and a combination of other available statistical software programs to perform statistical and

graphical analyses. Metric development was based

on a series of visual comparisons of community attribute responses to measures of disturbance (combination of selected water chemistry and land use

characteristics as discussed above) using box plots

and jittered dot density plots. Attributes that exhibited evidence of separation between reference

and impacted wetlands were selected as metrics

(or incorporated into the existing list of metrics).

Attributes that exhibited inconsistent or overlapping

responses between impacted and reference systems

were eliminated from further consideration.
90

 Lessons Learned

 Finding willing and cooperative private property landowners for access to potentially impacted wetlands is difficult!

 Determining what attribute or attributes (e.g.,

land use, chemical contaminant concentration,

distance to nearest road) to use as surrogate

measures of human disturbance is critical to the

successful development of reliable multimetric

biological indices.

 Drought (or floods) can seriously hamper sampling plans and interfere with the best of plans!

 The list of suitable metrics among macroinvertebrate communities of permanent wetlands is

different and much shorter than the list of metrics

suitable for wetlands with shorter water duration periods.
91

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Glossary

Aquatic life use A type of designated use pertaining to the support and maintenance of healthy biological communities.

Assemblage An association of interacting populations of organisms that belong to the same major

taxonomic groups. Examples of assemblages used

for bioassessments include: algae, amphibians, birds,

fish, amphibians, macroinvertebrates (insects, crayfish, clams, snails, etc.), and vascular plants.

Attribute A measurable component of a biological system. In the context of bioassessments, attributes include the ecological processes or characteristics of an individual or assemblage of species

that are expected, but not empirically shown, to

respond to a gradient of human disturbance.

Benthos The bottom fauna of waterbodies.

Biological assessment (bioassessment) Using

biomonitoring data of samples of living organisms

to evaluate the condition or health of a place (e.g.,

a stream, wetland, or woodlot).

Biological integrity “the ability of an aquatic ecosystem to support and maintain a balanced, adaptive community of organisms having a species composition, diversity, and functional organization comparable to that of natural habitats within a region”

(Karr and Dudley 1981).

Biological monitoring Sampling the biota of a

place (e.g., a stream, a woodlot, or a wetland).

Biota All the plants and animals inhabiting an area.

Composition (structure) The composition of the

taxonomic grouping such as fish, algae, or

macroinvertebrates relating primarily to the kinds

and number of organisms in the group.

Community All the groups of organisms living together in the same area, usually interacting or depending on each other for existence.

Competition Utilization by different species of limited resources of food or nutrients, refugia, space,

ovipositioning sites, or other resources necessary

for reproduction, growth, and survival.

Criteria A part of water quality standards. Criteria are the narrative and numeric definitions conditions that must be protected and maintained to support a designated use.

Continuum A gradient of change.

Designated use A part of water quality standards.

A designated use is the ecological goal that

policymakers set for a waterbody, such as aquatic

life use support, fishing, swimming, or drinking water.

Disturbance “Any discrete event in time that disrupts ecosystems, communities, or population structure and changes resources, substrate availability

or the physical environment” (Picket and White

1985). Examples of natural disturbances are fire,

drought, and floods. Human-caused disturbances

are referred to as “human disturbance” and tend to

be more persistent over time, e.g., plowing,

clearcutting of forests, conducting urban stormwater

into wetlands.

Diversity A combination of the number of taxa

(see taxa richness) and the relative abundance of

those taxa. A variety of diversity indexes have been

developed to calculate diversity.

Dominance The relative increase in the abundance

of one or more species in relation to the abundance

of other species in samples from a habitat.

Ecological risk assessment An evaluation of the

potential adverse effects that human activities have

on the plants and animals that make up ecosystems.

Ecosystem Any unit that includes all the organisms that function together in a given area interacting with the physical environment so that a flow of
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energy leads to clearly defined biotic structure and

cycling of materials between living and nonliving

parts (Odum 1983).

Ecoregion A region defined by similarity of climate, landform, soil, potential natural vegetation,

hydrology, and other ecologically relevant variables.

Gradient of human disturbance The relative

ranking of sample sites within a regional wetland

class based on degrees of human disturbance (e.g.,

pollution, physical alteration of habitats, etc.)

Habitat The sum of the physical, chemical, and

biological environment occupied by individuals of a

particular species, population, or community.

Hydrology The science of dealing with the properties, distribution, and circulation of water both on

the surface and under the earth.

Impact A change in the chemical, physical (including habitat), or biological quality or condition of a

waterbody caused by external forces.

Impairment Adverse changes occurring to an ecosystem or habitat. An impaired wetland has some

degree of human disturbance affecting it.

Index of biologic integrity (IBI) An integrative

expression of the biological condition that is composed of multiple metrics. Similar to economic indexes used for expressing the condition of the

economy.

Intolerant taxa Taxa that tend to decrease in wetlands or other habitats that have higher levels of

human disturbances, such as chemical pollution or

siltation.

Macroinvertebrates Animals without backbones

(insects, crayfish, clams, snails, etc.) that are caught

with a 500-800 micron mesh net.

Macroinvertebrates do not include zooplankton or

ostracods, which are generally smaller than 200 microns in size.

Metric An attribute with empirical change in value

along a gradient of human disturbance.

Minimally impaired site Sample sites within a

regional wetland class that exhibit the least degree

of detrimental effect. Such sites help anchor gradients of human disturbance and are commonly referred to as reference sites.

Most-impaired site Sample sites within a regional

wetland class that exhibit the greatest degree of

detrimental effect. Such sites help anchor gradients

of human disturbance and serve as important references, although they are not typically referred to as

reference sites.

Population A set of organisms belonging to the

same species and occupying a particular area at the

same time.

Reference site (as used with an index of biological integrity) A minimally impaired site that is

representative of the expected ecological conditions

and integrity of other sites of the same type and

region.

Stressor Any physical, chemical, or biological entity that can induce an adverse response.

Taxa A grouping of organisms given a formal taxonomic name such as species, genus, family, etc. The

singular form is taxon.

Taxa richness The number of distinct species or

taxa that are found in an assemblage, community,

or sample.

Tolerance The biological ability of different species or populations to survive successfully within a

certain range of environmental conditions.

Trophic Feeding, thus pertaining to energy

transfers.

Wetland(s) (1) Those areas that are inundated or

saturated by surface or groundwater at a frequency

and duration sufficient to support, and that under

normal circumstances do support, a prevalence of

vegetation typically adapted for life in saturated soil

conditions [EPA, 40 C.F.R.§ 230.3 (t) / USACE,

33 C.F.R. § 328.3 (b)]. (2) Wetlands are lands
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transitional between terrestrial and aquatic systems

where the water table is usually at or near the surface or the land is covered by shallow water. For

the purposes of this classification, wetlands must

have one or more of the following three attributes:

(a) at least periodically, the land supports predominantly hydrophytes, (b) the substrate is predominantly undrained hydric soil, and (c) the substrate is

nonsoil and is saturated with water or covered by

shallow water at some time during the growing season of each year (Cowardin et al. 1979). (3) The

term “wetland” except when such term is part of

the term “converted wetland,” means land that (a)

has a predominance of hydric soils, (b) is inundated

or saturated by surface or ground water at a frequency and duration sufficient to support a prevalence of hydrophytic vegetation typically adapted

for life in saturated soil conditions, and (c) under

normal circumstances does support a prevalence

of such vegetation. For purposes of this Act and

any other Act, this term shall not include lands in

Alaska identified as having a high potential for agricultural development which have a predominance

of permafrost soils [Food Security Act, 16 U.S.C.

801(a)(16)].